Compositions useful for treating disorders related to kit

ABSTRACT

Compounds and compositions useful for treating disorders related to Kit are described herein.

CLAIM OF PRIORITY

This application claims priority to U.S. Ser. No. 61/788,857 filed Mar.15, 2013, the contents of which is incorporated herein by reference inits entirety.

BACKGROUND

The invention relates to compounds and compositions useful for treatingdisorders related to Kit.

The enzyme Kit (also called CD117) is a receptor tyrosine kinase (RTK)expressed on a wide variety of cell types. The Kit molecule contains along extracellular domain, a transmembrane segment, and an intracellularportion. The ligand for Kit is stem cell factor (SCF), whose binding tothe extracellular domain of Kit induces receptor dimerization andactivation of downstream signaling pathways. Kit mutations generallyoccur in the DNA encoding the juxtumembrane domain (exon 11). They alsooccur, with less frequency, in exons 7, 8, 9, 13, 14, 17, and 18.Mutations make Kit function independent of activation by SCF, leading toa high cell division rate and possibly genomic instability. Mutant Kithas been implicated in the pathogenesis of several disorders andconditions including including systemic mastocytosis, GIST(gastrointestinal stromal tumors), AML (acute myeloid leukemia),melanoma, and seminoma. As such, there is a need for therapeutic agentsthat inhibit Kit, and especially agents that inhibit mutant Kit.

SUMMARY OF THE INVENTION

The present invention provides compounds and compositions for treatingor preventing conditions such as mastocytosis by modulating the activityof Kit, such compounds having the structural formula I:

or a pharmaceutically acceptable salt or a tautomer thereof, whereineach of the variables are described herein.

Any of the compounds disclosed herein may be used to treat any of thediseases disclosed herein.

DETAILED DESCRIPTION OF THE INVENTION

“Aliphatic group” means a straight-chain, branched-chain, or cyclichydrocarbon group and includes saturated and unsaturated groups, such asan alkyl group, an alkenyl group, and an alkynyl group.

“Alkenyl” means an aliphatic group containing at least one double bond.

“Alkoxyl” or “alkoxy” means an alkyl group having an oxygen radicalattached thereto. Representative alkoxyl groups include methoxy, ethoxy,propyloxy, tert-butoxy and the like.

“Alkyl” refers to a monovalent radical of a saturated straight orbranched hydrocarbon, such as a straight or branched group of 1-12,1-10, or 1-6 carbon atoms, referred to herein as C₁-C₁₂ alkyl, C₁-C₁₀alkyl, and C₁-C₆ alkyl, respectively. Exemplary alkyl groups include,but are not limited to, methyl, ethyl, propyl, isopropyl,2-methyl-1-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl,3-methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl,2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-1-pentyl,2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl,2,2-dimethyl-1-butyl, 3,3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl,isobutyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, heptyl, octyl,etc.

“Alkylene” refers to a divalent radical of an alkyl group.

“C₀ alkylene” refers to a bond. Thus, a moiety defined herein as“—(C₀-C₆ alkylene)-aryl” includes both -aryl (i.e., C₀ alkylene-aryl)and —(C₁-C₆ alkylene)-aryl.

“Halo” refers to a radical of any halogen, e.g., —F, —Cl, —Br, or —I.

“Haloalkyl” refers to an alkyl group that is substituted with at leastone halogen. For example, —CH₂F, —CHF₂, —CF₃, —CH₂CF₃, —CF₂CF₃, and thelike.

“Carbocyclic ring system” refers to a monocyclic, bicyclic or polycyclichydrocarbon ring system, wherein each ring is either completelysaturated or contains one or more units of unsaturation, but where noring is aromatic.

“Carbocyclyl” refers to a monovalent radical of a carbocyclic ringsystem. Representative carbocyclyl groups include cycloalkyl groups(e.g., cyclopentyl, cyclobutyl, cyclopentyl, cyclohexyl and the like),and cycloalkenyl groups (e.g., cyclopentenyl, cyclohexenyl,cyclopentadienyl, and the like).

“Aromatic ring system” is art-recognized and refers to a monocyclic,bicyclic or polycyclic hydrocarbon ring system, wherein at least onering is aromatic.

“Aryl” refers to a monovalent radical of an aromatic ring system.Representative aryl groups include fully aromatic ring systems, such asphenyl, naphthyl, and anthracenyl, and ring systems where an aromaticcarbon ring is fused to one or more non-aromatic carbon rings, such asindanyl, phthalimidyl, naphthimidyl, or tetrahydronaphthyl, and thelike.

“Heteroaromatic ring system” is art-recognized and refers to monocyclic,bicyclic or polycyclic ring system wherein at least one ring is botharomatic and comprises at least one heteroatom (e.g., N, O or S); andwherein no other rings are heterocyclyl (as defined below). In certaininstances, a ring which is aromatic and comprises a heteroatom contains1, 2, 3, or 4 ring heteroatoms in such ring.

“Heteroaryl” refers to a monovalent radical of a heteroaromatic ringsystem. Representative heteroaryl groups include ring systems where (i)each ring comprises a heteroatom and is aromatic, e.g., imidazolyl,oxazolyl, thiazolyl, triazolyl, pyrrolyl, furanyl, thiophenyl pyrazolyl,pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, indolizinyl, purinyl,naphthyridinyl, and pteridinyl; (ii) each ring is aromatic orcarbocyclyl, at least one aromatic ring comprises a heteroatom and atleast one other ring is a hydrocarbon ring or e.g., indolyl, isoindolyl,benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, carbazolyl, acridinyl, phenazinyl,phenothiazinyl, phenoxazinyl, pyrido[2,3-b]-1,4-oxazin-3(4H)-one,5,6,7,8-tetrahydroquinolinyl and 5,6,7,8-tetrahydroisoquinolinyl; and(iii) each ring is aromatic or carbocyclyl, and at least on aromaticring shares a bridgehead heteroatom with another aromatic ring, e.g.,4H-quinolizinyl.

“Heterocyclic ring system” refers to monocyclic, bicyclic and polycyclicring systems where at least one ring is saturated or partiallyunsaturated (but not aromatic) and comprises at least one heteroatom. Aheterocyclic ring system can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted.

“Heterocyclyl” refers to a monovalent radical of a heterocyclic ringsystem. Representative heterocyclyls include ring systems in which (i)every ring is non-aromatic and at least one ring comprises a heteroatom,e.g., tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, pyrrolidonyl,piperidinyl, pyrrolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl,dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl,and quinuclidinyl; (ii) at least one ring is non-aromatic and comprisesa heteroatom and at least one other ring is an aromatic carbon ring,e.g., 1,2,3,4-tetrahydroquinolinyl, 1,2,3,4-tetrahydroisoquinolinyl; and(iii) at least one ring is non-aromatic and comprises a heteroatom andat least one other ring is aromatic and comprises a heteroatom, e.g.,3,4-dihydro-1H-pyrano[4,3-c]pyridine, and1,2,3,4-tetrahydro-2,6-naphthyridine.

“Saturated heterocyclyl” refers to a radical of heterocyclic ring systemwherein every ring is saturated, e.g, tetrahydrofuran,tetrahydro-2H-pyran, pyrrolidine, piperidine and piperazine.

“Methylene” refers to the divalent radical —CH₂—.

“Ethane-1,2-diyl” refers to the divalent radical —CH₂CH₂—.

“Propane-1,3-diyl” refers to the divalent radical —CH₂CH₂CH₂—.

“Methylenedioxy” refers to the divalent radical —O—CH₂—O—.

“Substituted”, whether preceded by the term “optionally” or not, meansthat one or more hydrogens of the designated moiety are replaced with asuitable substituent. Unless otherwise indicated, an “optionallysubstituted” group may have a suitable substituent at each substitutableposition of the group, and when more than one position in any givenstructure may be substituted with more than one substituent selectedfrom a specified group, the substituent may be either the same ordifferent at each position. Combinations of substituents envisionedunder this invention are preferably those that result in the formationof stable or chemically feasible compounds. The term “stable”, as usedherein, refers to compounds that are not substantially altered whensubjected to conditions to allow for their production, detection, and,in certain embodiments, their recovery, purification, and use for one ormore of the purposes disclosed herein.

Suitable monovalent substituents on a substitutable carbon atom of an“optionally substituted” group (such as an alkyl, alkenyl, alkynyl,alkylene, alkenylene, alkynylene or the carbon atom of a carbocyclyl,aryl, heterocyclyl or heteroaryl) are independently halogen;—(CH₂)₀₋₄R^(∘); —(CH₂)₀₋₄OR^(∘); —O—(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄CH(OR^(∘))₂; —(CH₂)₀₋₄SR^(∘); —(CH₂)₀₋₄Ph, which may besubstituted with R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁Ph which may be substitutedwith R^(∘); —CH═CHPh, which may be substituted with R^(∘); —NO₂; —CN;—N₃; —(CH₂)₀₋₄N(R^(∘))₂; —(CH₂)₀₋₄N(R^(∘))C(O)R^(∘); —N(R^(∘))C(S)R^(∘);—(CH₂)₀₋₄N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))C(S)NR^(∘) ₂;—(CH₂)₀₋₄N(R^(∘))C(O)OR^(∘); —N(R^(∘))N(R^(∘))C(O)R^(∘);—N(R^(∘))N(R^(∘))C(O)NR^(∘) ₂; —N(R^(∘))N(R^(∘))C(O)OR^(∘);—(CH₂)₀₋₄C(O)R^(∘); —C(S)R^(∘); —(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄C(O)SR^(∘); —(CH₂)₀₋₄C(O)OSiR^(∘) ₃; —(CH₂)₀₋₄OC(O)R^(∘);—OC(O)(CH₂)₀₋₄SR^(∘)—, SC(S)SR^(∘); —(CH₂)₀₋₄SC(O)R^(∘);—(CH₂)₀₋₄C(O)NR^(∘) ₂; —C(S)NR^(∘) ₂; —C(S)SR^(∘); —(CH₂)₀₋₄OC(O)NR^(∘)₂; —C(O)N(OR^(∘))R^(∘); —C(O)C(O)R^(∘); —C(O)CH₂C(O)R^(∘);—C(NOR^(∘))R^(∘); —(CH₂)₀₋₄SSR^(∘); —(CH₂)₀₋₄S(O)₂R^(∘);—(CH₂)₀₋₄S(O)₂OR^(∘); —(CH₂)₀₋₄OS(O)₂R^(∘); —S(O)₂NR^(∘) ₂;—(CH₂)₀₋₄S(O)R^(∘); —N(R^(∘))S(O)₂NR^(∘) ₂; —N(R^(∘))S(O)₂R^(∘);—C(O)—N(R^(∘))—S(O)₂—R^(∘); —N(OR^(∘))R^(∘); —C(NH)NR^(∘) ₂;—P(O)₂R^(∘); —P(O)R^(∘) ₂; —OP(O)R^(∘) ₂; —OP(O)(OR^(∘))₂; —SiR^(∘) ₃;—(C₁₋₄ straight or branched)alkylene)O—N(R^(∘))₂; or —(C₁₋₄ straight orbranched)alkylene)C(O)O—N(R^(∘))₂, wherein each R^(∘) may be substitutedas defined below and is independently hydrogen, C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur, or, notwithstanding the definition above, twoindependent occurrences of R^(∘), taken together with their interveningatom(s), form a 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur, which may be substituted as definedbelow.

Suitable monovalent substituents on R^(∘) (or the ring formed by takingtwo independent occurrences of R^(∘) together with their interveningatoms), are independently halogen, —(CH₂)₀₋₂R^(), -(haloR^()),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(), —(CH₂)₀₋₂CH(OR^())₂; —O(haloR^()), —CN,—N₃, —(CH₂)₀₋₂C(O)R^(), —(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(),—(CH₂)₀₋₂SR^(), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NHR^(),—(CH₂)₀₋₂NR^() ₂, —NO₂, —OSiR^() ₃, —C(O)SR^(), —(C₁₋₄ straight orbranched alkylene)C(O)OR^(), or —SSR^() wherein each R^() isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently selected from C₁₋₄ aliphatic,—CH₂Ph, —O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents on asaturated carbon atom of R^(∘) include ═O and ═S.

Suitable divalent substituents on a saturated carbon atom of an“optionally substituted” group include the following: ═O (“oxo”), ═S,═NNR*₂, ═NNHC(O)R*, ═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*,—O(C(R*₂))₂₋₃O—, or —S(C(R*₂))₂₋₃S—, wherein each independent occurrenceof R* is selected from hydrogen, C₁₋₆ aliphatic which may be substitutedas defined below, or an unsubstituted 5-6-membered saturated, partiallyunsaturated, or aryl ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur. Suitable divalent substituents thatare bound to vicinal substitutable carbons of an “optionallysubstituted” group include: —O(CR*₂)₂₋₃O—, wherein each independentoccurrence of R* is selected from hydrogen, C₁₋₆ aliphatic which may besubstituted as defined below, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R* include halogen,—R^()-(haloR^()), —OH, —OR^(), —O(haloR^()), —CN, —C(O)OH,—C(O)OR^(), —NH₂, —NHR^(), —NR^() ₂, or —NO₂, wherein each R^() isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

Suitable substituents on a substitutable nitrogen of an “optionallysubstituted” group include —R^(†), —NR^(†) ₂, —C(O)R^(†), —C(O)OR^(†),—C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†), —S(O)₂R^(†), —S(O)₂NR^(†) ₂,—C(S)NR^(†) ₂, —C(NH)NR^(†) ₂, or —N(R^(†))S(O)₂R^(†); wherein eachR^(†) is independently hydrogen, C₁₋₆ aliphatic which may be substitutedas defined below, unsubstituted —OPh, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(†), taken together with their intervening atom(s) form anunsubstituted 3-12-membered saturated, partially unsaturated, or arylmono- or bicyclic ring having 0-4 heteroatoms independently selectedfrom nitrogen, oxygen, or sulfur.

Suitable substituents on the aliphatic group of R^(†) are independentlyhalogen, —R^(†), -(haloR^(†)), —OH, —OR*, —O(haloR^(†)), —CN, —C(O)OH,—C(O)OR^(†), —NH₂, —NHR^(†), —NR^(†) ₂, or —NO₂, wherein each R^(†) isunsubstituted or where preceded by “halo” is substituted only with oneor more halogens, and is independently C₁₋₄aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

As used herein, the definition of each expression, e.g., alkyl, m, n,etc., when it occurs more than once in any structure, is intended to beindependent of its definition elsewhere in the same structure.

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the invention. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in this invention.

If, for instance, a particular enantiomer of compound of the presentinvention is desired, it may be prepared by asymmetric synthesis, or byderivation with a chiral auxiliary, where the resulting diastereomericmixture is separated and the auxiliary group cleaved to provide the puredesired enantiomers. Alternatively, where the molecule contains a basicfunctional group, such as amino, or an acidic functional group, such ascarboxyl, diastereomeric salts are formed with an appropriateoptically-active acid or base, followed by resolution of thediastereomers thus formed by fractional crystallization orchromatographic means well known in the art, and subsequent recovery ofthe pure enantiomers.

Unless otherwise indicated, when a disclosed compound is named ordepicted by a structure without specifying the stereochemistry and hasone or more chiral centers, it is understood to represent all possiblestereoisomers of the compound, as well as enantiomeric mixtures thereof.

The “enantiomeric excess” or “% enantiomeric excess” of a compositioncan be calculated using the equation shown below. In the example shownbelow a composition contains 90% of one enantiomer, e.g., the Senantiomer, and 10% of the other enantiomer, i.e., the R enantiomer.

ee=(90−10)/100=80%.

Thus, a composition containing 90% of one enantiomer and 10% of theother enantiomer is said to have an enantiomeric excess of 80%.

The compounds or compositions described herein may contain anenantiomeric excess of at least 50%, 75%, 90%, 95%, or 99% of one formof the compound, e.g., the S-enantiomer. In other words such compoundsor compositions contain an enantiomeric excess of the S enantiomer overthe R enantiomer.

The compounds described herein may also contain unnatural proportions ofatomic isotopes at one or more of the atoms that constitute suchcompounds. For example, the compounds may be radiolabeled withradioactive isotopes, such as for example deuterium (²H), tritium (³H),carbon-13 (¹³C), or carbon-14 (¹⁴C). All isotopic variations of thecompounds disclosed herein, whether radioactive or not, are intended tobe encompassed within the scope of the present invention. In addition,all tautomeric forms of the compounds described herein are intended tobe within the scope of the invention.

The compound can be useful as the free base or as a salt. Representativesalts include the hydrobromide, hydrochloride, sulfate, bisulfate,phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate,laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate,fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate,lactobionate, and laurylsulphonate salts and the like. (See, forexample, Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm. Sci.66:1-19.)

Certain compounds disclosed herein can exist in unsolvated forms as wellas solvated forms, including hydrated forms. In general, the solvatedforms are equivalent to unsolvated forms and are encompassed within thescope of the present invention. Certain compounds disclosed herein mayexist in multiple crystalline or amorphous forms. In general, allphysical forms are equivalent for the uses contemplated by the presentinvention and are intended to be within the scope of the presentinvention.

As used herein, the term “patient” refers to organisms to be treated bythe methods of the present invention. Such organisms preferably include,but are not limited to, mammals (e.g., murines, simians, equines,bovines, porcines, canines, felines, and the like), and most preferablyincludes humans.

As used herein, the term “effective amount” refers to the amount of acompound (e.g., a compound of the present invention) sufficient toeffect beneficial or desired results. An effective amount can beadministered in one or more administrations, applications or dosages andis not intended to be limited to a particular formulation oradministration route. As used herein, the term “treating” includes anyeffect, e.g., lessening, reducing, modulating, ameliorating oreliminating, that results in the improvement of the condition, disease,disorder, and the like, or ameliorating a symptom thereof.

Compounds

In one embodiment, the invention provides a compound having structuralformula I:

or a pharmaceutically acceptable salt or a tautomer thereof, wherein:

each R⁴ is independently selected from —C(O)N(R²)(R²), —C₁-C₄ alkyl,—CN, —(C₀-C₄ alkylene)-N(R²)(R²), —O—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl),—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl), or —O—(C₀-C₄ alkylene)-(R¹), whereinR¹ is selected from —C₁-C₄ alkyl and heterocyclyl;

each R² is independently selected from hydrogen and unsubstituted C₁-C₄alkyl

each R⁶ is independently selected from —C₁-C₄ alkyl or two R⁶ bound tothe same carbon atom are taken together to form oxo, or two R⁶ bound todifferent carbon atoms are taken together to form methylene,ethane-1,2-diyl, or propane-1,2-diyl forming a ring that is bridged tothe piperazine-1,4-diyl portion of the compound;

each R⁸ is independently selected from halo, —OH, —N(R²)(R²), C₁-C₄alkyl, and —O—(C₁-C₄ alkyl);

L is selected from a bond, —(C₁-C₄ alkylene)-, —O—, —S—, —SO₂—, —N(R²)—,—O—(C₁-C₄ alkylene)-, —(C₁-C₄ alkylene)-O—, —(C₁-C₄ alkylene)-N(R²)—,—N(R²)—(C₁-C₄ alkylene)-, —N(R²)—CO—(C₁-C₄ alkylene)-, —CO—N(R²)—(C₁-C₄alkylene)-;

n is 0, 1, 2, or 3;

m is 0, 1, 2, 3, or 4;

p is 0, 1, or 2;

q is 0, 1, 2, or 3;

ring A is monocyclic or bicyclic aryl, heteroaryl, carbocyclyl orheterocyclyl;

each R¹⁰ is independently selected from halo; —OH, —CN, —C(O)N(R²)(R²),C₁-C₄ alkyl, —O—(C₁-C₄ alkyl), or heterocyclyl, or two R¹° bound toadjacent ring carbon atoms are taken together to form methylenedioxy;

wherein unless otherwise specified any alkyl, or alkylene portion of thecompound is optionally substituted.

In some embodiments, the compound of Formula I is not

In certain embodiments of the compound of Formula I, any alkyl, oralkylene portion of the compound is optionally and independentlysubstituted with one or more substituents independently selected fromhalo, —OH, C₁-C₄ alkyl, C₁-C₄ haloalkyl, —O—(C₁-C₄ alkyl), or ═O; andany heterocyclyl portion of the compound is optionally and independentlysubstituted with one or more substituents independently selected fromhalo, —OH, C₁-C₄ alkyl, C₁-C₄ haloalkyl, —O—(C₁-C₄ alkyl), or ═O.

In certain embodiments of the compound of Formula I, each R⁴ isindependently selected from —C(O)N(R²)(R²), —O—(C₁-C₄ alkylene)-O—(C₁-C₃alkyl), and —O—(C₀-C₄ alkylene)-(R¹), wherein:

R¹ is selected from —C₁-C₄ alkyl, and a saturated heterocyclyl;

each R² is independently selected from hydrogen and methyl;

each alkyl or alkylene is optionally substituted with one or moresubstituents independently selected from halo, —OH and —O(C₁-C₃ alkyl);

the saturated heterocyclyl is optionally substituted on a substitutablering nitrogen with methyl; and

the saturated heterocyclyl is optionally substituted on a substitutablering carbon with with one or more substituents independently selectedfrom methyl, halo, —OH.

In one aspect of the above embodiments, each R⁴ is independentlyselected from C(O)NH₂, —OCF₂, —OCH₂CH₃, —OCH₃,1-methyl-4-fluoropiperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-yloxy,1-(2-hydroxyethyl)-pyrrolidin-3-yloxy, 1-methylazetidin-3-ylmethoxy,1-methylpiperidin-3-ylethoxy, 1-methylpiperidin-3-ylmethoxy,1-methylpiperidin-4-ylmethoxy, 1-methylpiperidin-4-yloxy,1-methylpyrrolidin-3-ylmethoxy, 1-methylpyrrolidin-3-yloxy,2,3-dihydroxypropoxy, 2-oxopiperidin-4-ylmethoxy,2-oxopyrrolidin-1-ylethoxy, 3-methyloxetan-3-ylmethoxy,4-methylmorpholin-2-ylmethoxy, 4-methylpiperazin-1-ylethoxy,4-methylpiperazin-1-ylpropoxy, azetidin-3-ylmethoxy, methoxyethoxy,morpholin-2-ylmethoxy, piperidin-1-ylpropoxy, piperidin-3-ylmethoxy,piperidin-4-ylmethoxy, piperidin-4-yloxy, pyrrolidin-1-ylpropoxy,pyrrolidin-3-ylmethoxy, and pyrrolidin-3-yloxy.

In a more specific aspect of these embodiments, each R⁴ is independentlyselected from OCH₂CH₃, OCH₃, 1-(2-hydroxyethyl)-piperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-yloxy,1-(2-hydroxyethyl)-pyrrolidin-3-yloxy,1-methyl-4-fluoropiperidin-4-ylmethoxy, 1-methylazetidin-3-ylmethoxy,1-methylpiperidin-3-ylethoxy, 1-methylpiperidin-3-ylmethoxy,1-methylpiperidin-4-ylmethoxy, 1-methylpiperidin-4-yloxy,1-methylpyrrolidin-3-ylmethoxy, 1-methylpyrrolidin-3-yloxy,2,3-dihydroxypropoxy, 2-oxopiperidin-4-ylmethoxy,2-oxopyrrolidin-1-yleithoxy, 3-methyloxetan-3-ylmethoxy,4-methylmorpholin-2-ylmethoxy, 4-methylpiperazin-1-ylethoxy,4-methylpiperazin-1-ylpropoxy, azetidin-3-ylmethoxy, methoxyethoxy,morpholin-2-ylmethoxy, piperidin-1-ylpropoxy, piperidin-3-ylmethoxy,piperidin-4-ylmethoxy, piperidin-4-yloxy, pyrrolidin-3-ylmethoxy, andpyrrolidin-3-yloxy.

In some embodiments of the compound of Formula I, each R⁶ isindependently selected from C₁-C₄ alkyl, ═O, or two R⁶ on non-adjacentcarbon ring atoms are taken together to form methylene, ethane-1,2-diyl,or propane-1,3-diyl thereby forming a ring that is bridged to thepiperazine-1,4-diyl portion of the compound. In the interest of clarity,examples of the bridged rings so formed are depicted below:

Two R⁶ form methylene:

Two R⁶ form ethane-1,2-diyl:

Two R⁶ form propane-1,3-diyl:

In a more specific aspect of the above embodiments, each R⁶ isindependently methyl or ═O, or two R⁶ non-adjacent carbon ring atoms aretaken together to form an ethane-1,2-diyl thereby forming a ring that isbridged to the piperazine-1,4-diyl portion of the compound.

In certain embodiments of the compound of Formula I each R⁸ isindependently selected from ═O and —NH₂.

In certain embodiments of the compound of Formula I each R¹⁰ isindependently selected from OH, —OCH₃, —F, —CH₃, —CN, —C(O)NH₂, —OCF₂,and —Cl. In certain embodiments of the compound of Formula I, L isselected from a bond, —CH₂—, —CH₂CH₂, NH, O, S, CH₂O—*, —OCH₂—*,—OCH(CH₃)—*, —N(CH₃)CH₂—*, —NHCH₂—*, —NHC(O)CH₂—*, —C(O)NH—*,—NHCH(CH₃)—*, and —SO₂—, wherein “*” represents a portion of L bound toring A. In one aspect of these embodiments, L is selected from —CH₂—,—CH₂CH₂—, —NH—, —O—, —S—, —CH₂O—*, —N(CH₃)CH₂—*, —OCH(CH₃)—*, and—OCH(CH₃)—*. In a more specific aspect of these embodiments, L is —CH₂—.

In certain embodiments of the compound of Formula I, ring A is selectedfrom phenyl, thiophenyl, indolinyl, 1,2,3,4-tetrahydroquinoline,pyridinyl, thiophenyl, and C₃-C₆ cycloalkyl. In one aspect of theseembodiments, ring A is selected from phenyl, and thiophen-2-yl. In amore specific aspect, ring A is phenyl.

In certain embodiment, the compound of the invention has structuralformula II:

or a pharmaceutically acceptable salt or tautomer thereof, wherein:

one of R^(4a) or R^(4b) is selected from hydrogen, —O—CH₃, —O—CH₂CH₃ and—O—CH₂CH₂—O—CH₃;

the other of R^(4a) or R^(4b) is selected from —O—CH₃, —O—CH₂CH₃,—O—CH₂CH₂—O—CH₃ and —(C₀-C₄ alkylene)-(saturated heterocyclyl), wherein:

the saturated heterocyclyl is optionally substituted on a substitutablering nitrogen with methyl; and

the saturated heterocyclyl is optionally substituted on a substitutablering carbon with one or more substituents independently selected frommethyl, halo, and —OH;

R^(6a) is hydrogen;

R^(6b) is selected from hydrogen, ═O, and —CH₃; or

R^(6a) and R^(6b) are taken together with the carbon atoms to which theyare bound to form a cyclobutyl fused to the piperazine-1,4-diyl ring;

R^(8a) is selected from hydrogen, —NH₂ and —OH;

L is selected from —CH₂—, —CH₂CH₂—, —NH—, —O—, —S—, —CH₂O—*,—N(CH₃)CH₂—*, —OCH(CH₃)—*, and —OCH(CH₃)—*;

R^(10a) is selected from hydrogen, —OCH₃, —C(O)NH₂ and halo; and

R^(10b) is selected from hydrogen, —OCH₃, —OCF₂, halo, —CH₃, CN, and OH.

In some embodiments of Formula II:

one of R^(4a) or R^(4b) is selected from hydrogen, —O—CH₃, —O—CH₂CH₃ and—O—CH₂CH₂—O—CH₃;

the other of R^(4a) or R^(4b) is selected from —O—CH₃, —O—CH₂CH₃,—O—CH₂CH₂—O—CH₃, 1-(2-hydroxyethyl)-piperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-yloxy,1-(2-hydroxyethyl)-pyrrolidin-3-yloxy,1-methyl-4-fluoropiperidin-4-ylmethoxy, 1-methylazetidin-3-ylmethoxy,1-methylpiperidin-3-ylethoxy, 1-methylpiperidin-3-ylmethoxy,1-methylpiperidin-4-ylmethoxy, 1-methylpiperidin-4-yloxy,1-methylpyrrolidin-3-ylmethoxy, 1-methylpyrrolidin-3-yloxy,2,3-dihydroxypropoxy, 2-oxopiperidin-4-ylmethoxy,2-oxopyrrolidin-1-yleithoxy, 3-methyloxetan-3-ylmethoxy,4-methylmorpholin-2-ylmethoxy, 4-methylpiperazin-1-ylethoxy,4-methylpiperazin-1-ylpropoxy, azetidin-3-ylmethoxy,morpholin-2-ylmethoxy, piperidin-1-ylpropoxy, piperidin-3-ylmethoxy,piperidin-4-ylmethoxy, piperidin-4-yloxy, pyrrolidin-3-ylmethoxy, andpyrrolidin-3-yloxy;

R^(10a) is selected from hydrogen, —OCH₃, and fluoro; and

R^(10b) is selected from hydrogen, —OCH₃, —OCF₂, fluoro, chloro, —CH₃,CN, OH, and —C(O)NH₂.

In still another embodiment, the invention provides a compound havingstructural formula III:

or a pharmaceutically acceptable salt or tautomer thereof, wherein

each of X, Y, and Z is independently, C or N, each of which may beattached to one or more R⁴;

each R⁴ is independently selected from —C(O)N(R²)(R²), —C₁-C₄ alkyl,—CN, —(C₀-C₄ alkylene)-N(R²)(R²), —O—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl),—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl), or —O—(C₀-C₄ alkylene)-(R¹), whereinR¹ is selected from —C₁-C₄ alkyl and heterocyclyl;

each R² is independently selected from hydrogen and unsubstituted C₁-C₄alkyl;

each R⁶ is independently selected from —C₁-C₄ alkyl or two R⁶ bound tothe same carbon atom are taken together to form oxo, or two R⁶ bound todifferent carbon atoms are taken together to form methylene,ethane-1,2-diyl or propane-1,3-diyl thereby forming a ring that isbridged to the piperazine-1,4-diyl portion of the compound;

each R⁸ is independently selected from halo, —OH, —N(R²)(R²), C₁-C₄alkyl, and —O—(C₁-C₄ alkyl);

L is selected from a bond, —(C₁-C₄ alkylene)-, —O—, —S—, —SO₂—, —N(R²)—,—O—(C₁-C₄ alkylene)-, —(C₁-C₄ alkylene)-O—, —(C₁-C₄ alkylene)-N(R²)—,—N(R²)—(C₁-C₄ alkylene)-, —N(R²)—CO—(C₁-C₄ alkylene)-, —CO—N(R²)— (C₁-C₄alkylene)-;

n is 0, 1, 2, or 3;

m is 0, 1, 2, 3, or 4;

p is 0, 1, or 2;

q is 0, 1, 2, or 3;

ring A is monocyclic or bicyclic aryl, heteroaryl, carbocyclyl orheterocyclyl;

each R¹⁰ is independently selected from halo; —OH, —CN, —C(O)N(R²)(R²),C₁-C₄ alkyl, —O—(C₁-C₄ alkyl), or heterocyclyl, or two R¹° bound toadjacent ring carbon atoms are taken together to form methylenedioxy;and

wherein unless otherwise specified any alkyl, or alkylene portion of thecompound is optionally substituted.

Indications

The compounds described herein can be useful for treating conditionsassociated with aberrant Kit activity. Activating mutations in Kit arefound in multiple indications, including systemic mastocytosis, GIST(gastrointestinal stromal tumors), AML (acute myeloid leukemia),melanoma, and seminoma.

Mastocytosis refers to a group of disorders characterized by excessivemast cell accumulation in one tissue, or in multiple tissues.Mastocytosis is subdivided into two groups of disorders: (1) cutaneousmastocytosis (CM) describes forms that are limited to the skin; and (2)systemic mastocytosis (SM) describes forms in which mast cellsinfiltrate extracutaneous organs, with or without skin involvement. SMis further subdivided into four forms: indolent (ISM), smoldering (SSM),aggressive (ASM), SM with associated hemotologic non-mast cell lineagedisease (SM-AHNMD), and mast cell leukemia (MCL).

Diagnosis of systemic mastocytosis is based on histological andcytological study of bone marrow showing infiltration by mast cells offrequently atypical morphology, which frequently abnormally expressnon-mast cell markers (CD25 and/or CD2). An elevated level of serumtryptase above 20 ng/mL or the presence of an activating mutation of Kitcan confirm the diagnosis. The extent of the proliferation of mast cellscan be analyzed using medical imaging (radiography, ultrasound, CTscanning).

Activating mutations at the D816 position are found in the vast majorityof mastocytosis cases, with the most common mutations being D816V andD816H, and D816Y. The D816V mutation is found in the activation loop ofthe kinase domain, and leads to constitutive activation.

The compounds described herein may also be useful to treat GIST.Complete surgical resection remains the principal treatment of choicefor patients with a primary GIST. Surgery is effective in approximately50% of patients with GIST. Of the remaining patients, tumor recurrenceis frequent. Primary treatment with a Kit inhibitor such as imatinib hasalso been shown to be sufficient for initial treatment. However,resistance to imatinib occurs within months through somatic mutation.These secondary imatinib resistant mutations are most frequently locatedon exon 11, 13, 14, 17 or 18. Sunitinib is the standard of care secondline treatment for most imatinib resistant tumors and is effective forthose containing mutations in exons 11, 13 and 14. However, secondarykit mutations in exons 17 and 18 are resistant to sunitinib treatmentand furthermore, tumors containing tertiary resistance mutations in exon17 and 18 emerge several months after sunitinib treatment. Regorafenibhas shown promising results in a phase 3 clinical trial of imatinib,sunitinib resistant GISTs with activity against several exon 17 and 18mutations, with the exception of the D816 mutant which remainsrefractory to all treatments. Regorafenib has been approved for 3rd lineGIST treatment.

The compounds described herein may also be useful in treating AML. SomeAML patients harbor Kit mutations as well, with the majority of thesemutations at the D816 position.

In addition, mutations in Kit have been linked to Ewing's sarcoma, DLBCL(diffuse large B cell lymphoma), dysgerminoma, MDS (myelodysplasticsyndrome), NKTCL (nasal NK/T-cell lymphoma), and CMML (chronicmyelomonocytic leukemia).

Pharmaceutical Compositions

While it is possible for a compound disclosed herein to be administeredalone, it is preferable to administer the compound as a pharmaceuticalformulation, where the compound is combined with one or morepharmaceutically acceptable excipients or carriers. The compoundsdisclosed herein may be formulated for administration in any convenientway for use in human or veterinary medicine. In certain embodiments, thecompound included in the pharmaceutical preparation may be activeitself, or may be a prodrug, e.g., capable of being converted to anactive compound in a physiological setting.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

Examples of pharmaceutically acceptable carriers include: (1) sugars,such as lactose, glucose and sucrose; (2) starches, such as corn starchand potato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; (21)cyclodextrins such as Captisol®; and (22) other non-toxic compatiblesubstances employed in pharmaceutical formulations.

Examples of pharmaceutically acceptable antioxidants include: (1) watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Solid dosage forms (e.g., capsules, tablets, pills, dragees, powders,granules and the like) can include one or more pharmaceuticallyacceptable carriers, such as sodium citrate or dicalcium phosphate,and/or any of the following: (1) fillers or extenders, such as starches,lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders,such as, for example, carboxymethylcellulose, alginates, gelatin,polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants, such asglycerol; (4) disintegrating agents, such as agar-agar, calciumcarbonate, potato or tapioca starch, alginic acid, certain silicates,and sodium carbonate; (5) solution retarding agents, such as paraffin;(6) absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as, for example, cetyl alcohol and glycerolmonostearate; (8) absorbents, such as kaolin and bentonite clay; (9)lubricants, such a talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and(10) coloring agents.

Liquid dosage forms can include pharmaceutically acceptable emulsions,microemulsions, solutions, suspensions, syrups and elixirs. In additionto the active ingredient, the liquid dosage forms may contain inertdiluents commonly used in the art, such as, for example, water or othersolvents, solubilizing agents and emulsifiers, such as ethyl alcohol,isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol,benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (inparticular, cottonseed, groundnut, corn, germ, olive, castor and sesameoils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Ointments, pastes, creams and gels may contain, in addition to an activecompound, excipients, such as animal and vegetable fats, oils, waxes,paraffins, starch, tragacanth, cellulose derivatives, polyethyleneglycols, silicones, bentonites, silicic acid, talc and zinc oxide, ormixtures thereof.

Powders and sprays can contain, in addition to an active compound,excipients such as lactose, talc, silicic acid, aluminum hydroxide,calcium silicates and polyamide powder, or mixtures of these substances.Sprays can additionally contain customary propellants, such aschlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, suchas butane and propane.

The formulations may conveniently be presented in unit dosage form andmay be prepared by any methods well known in the art of pharmacy. Theamount of active ingredient which can be combined with a carriermaterial to produce a single dosage form will vary depending upon thehost being treated, the particular mode of administration. The amount ofactive ingredient that can be combined with a carrier material toproduce a single dosage form will generally be that amount of thecompound which produces a therapeutic effect.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically acceptablecarrier, and with any preservatives, buffers, or propellants that may berequired.

When the compounds disclosed herein are administered as pharmaceuticals,to humans and animals, they can be given per se or as a pharmaceuticalcomposition containing, for example, 0.1 to 99.5% (more preferably, 0.5to 90%) of active ingredient in combination with a pharmaceuticallyacceptable carrier.

The formulations can be administered topically, orally, transdermally,rectally, vaginally, parentally, intranasally, intrapulmonary,intraocularly, intravenously, intramuscularly, intraarterially,intrathecally, intracapsularly, intradermally, intraperitoneally,subcutaneously, subcuticularly, or by inhalation.

Dosages

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound disclosed hereinemployed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compound employed, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound that is the lowest dose effective to producea therapeutic effect. Such an effective dose will generally depend uponthe factors described above. Generally, intravenous,intracerebroventricular and subcutaneous doses of the compounds of thisinvention for a patient will range from about 0.0001 to about 100 mg perkilogram of body weight per day. If desired, the effective daily dose ofthe active compound may be administered as two, three, four, five, sixor more sub-doses administered separately at appropriate intervalsthroughout the day, optionally, in unit dosage forms. In someembodiments, the dose will be 1-20, or 5-10 mg per kilogram of bodyweight, administered twice daily.

EXAMPLES

The following examples are intended to be illustrative, and are notmeant in any way to be limiting.

Example 1

Step 1:

In a 75 mL sealed vessel, 4-chloro-6,7-dimethoxyquinazoline (1 g, 4.45mmol) and 5-bromo-2-(piperazin-1-yl)pyrimidine (1.19 g, 4.89 mmol) weredissolved in isopropanol (30 mL) and triethylamine (0.745 mL, 5.34mmol). The reaction mixture was stirred at 90 degrees Celsius for 15hours. The mixture was then diluted with water and the resulting solidswere filtered, washed with water and dried to give the desired product4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline asa white solid (1.85 g). This material was used in Step 2 without furtherpurification.

Step 2:

In a sealable reaction tube,4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline (50mg, 0.116 mmol), 2-benzyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.031mL, 0.14 mmol), potassium carbonate (48 mg, 0.35 mmol) and Pd(PPh₃)₄(13.5 mg, 0.0116 mmol), were combined with 1/1 THF and water (1.3 mL) ina N₂ atmosphere and heated to 100 degrees Celsius for 15 hours. Thecooled reaction mixture was diluted with ethyl acetate and filteredthrough a celite plug. The residue was purified by silica gelchromatography (0-10% methanol/methylene chloride) followed bypreparative thin layer chromatography eluting with 1/1 acetone indichloromethane. The desired product was excised and extracted with 10%methanol in dichloromethane. After drying on high vacuum, the product4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazolinewas obtained as an off-white solid (25 mg, 49% yield).

Example 2

Step 1:

In a sealable reaction tube,4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline(0.4 g, 0.93 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (0.425 g,1.8 mmol), potassium acetate (0.46 g, 4.65 mmol) and[1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II), complexwith dichloromethane (38 mg, 0.0465 mmol), were combined in anhydrousDMSO (4.65 mL) in a N₂ atmosphere and heated to 80 degrees Celsius for15 hours. The mixture was then diluted with water and the resultingsolids were filtered, washed with water and dried to give the desiredproduct6,7-dimethoxy-4-(4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-yl)piperazin-1-yl)quinazolineas a pale brown solid (0.417 g).

Step 2:

In a sealable reaction tube,6,7-dimethoxy-4-(4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-yl)piperazin-1-yl)quinazoline(35 mg, 0.073 mmol), 1-(bromomethyl)-4-fluorobenzene (0.011 mL, 0.088mmol), cesium carbonate (71 mg, 0.219 mmol) and Pd(^(t)Bu₃)₃ (5 mg,0.009 mmol), were combined in 2/1 THF and water (1.2 mL) in a N₂atmosphere and heated to 140 degrees Celsius for 15 minutes in a MWreactor. The cooled reaction mixture was diluted with ethyl acetate andfiltered through a celite plug. The residue was purified by silica gelchromatography (0-6% methanol/methylene chloride). After drying on highvacuum, the product4-(4-(5-(4-fluorobenzyl)pyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazolinewas obtained as an off-white solid (21 mg, 64% yield).

Example 3

In a sealable reaction tube,4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline (50mg, 0.12 mmol), benzyl alcohol (0.12 mL, 1.163 mmol), copper iodide (2.2mg, 0.012 mmol), 1,10-phenanthroline (4.2 mg, 0.023 mmol), and cesiumcarbonate (57 mg, 0.174 mmol), were combined in anhydrous toluene (0.5mL) in a N₂ atmosphere and heated to 110 degrees Celsius for 48 hours.The cooled reaction mixture was diluted with ethyl acetate and filteredthrough a celite plug. The residue was purified by silica gelchromatography (0-6% methanol/methylene chloride. After drying on highvacuum, the product4-(4-(5-(benzyloxy)pyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazolinewas obtained as an off-white solid (37 mg, 67% yield).

Example 4

Step 1:

In a vessel, tert-butyl4-(5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-yl)piperazine-1-carboxylate(0.5 g, 1.28 mmol) and 35% aqueous hydrogen peroxide (1.2 mL, 12.8 mmol)were dissolved in tetrahydrofuran (10 mL). The reaction mixture wasstirred at room temperature for 1.5 hours. The mixture diluted to 40 mLwith aqueous saturated sodium thiosulfate solution and then extractedwith ethylacetate to give the desired product tert-butyl4-(5-hydroxypyrimidin-2-yl)piperazine-1-carboxylate as a brown oil. Thismaterial was used in Step 2 without further purification.

Step 2:

In a vessel, tert-butyl4-(5-hydroxypyrimidin-2-yl)piperazine-1-carboxylate from the previousstep (1.28 mmol) and 4M hydrogen chloric acid in dioxane (4 mL, 12.8mmol) were dissolved in 2/1 mixture of methanol and dichloromethane (10mL). The reaction mixture was stirred at room temperature for 1.5 hours.The mixture was then concentrated to dryness to give the desired product2-(piperazin-1-yl)pyrimidin-5-ol as a brown solid. This material wasused in Step 3 without further purification.

Step 3:

In a microwave vessel, 2-(piperazin-1-yl)pyrimidin-5-ol from theprevious step (1.28 mmol) and 4-chloro-6,7-dimethoxyquinazoline (0.29 g,1.28 mmol) were dissolved in isopropanol (6 mL) and diisopropylethylamine (0.23 mL, 1.41 mmol). The reaction mixture was heated at 140degrees Celsius in a microwave reactor for 30 minutes. The mixture wasthen diluted with hexanes and the resulting solids were filtered, washedwith hexanes and dried to give the desired product2-(4-(6,7-dimethoxyquinazolin-4-yl)piperazin-1-yl)pyrimidin-5-ol as abrown solid (0.42 g). This material was used in the next step withoutfurther purification.

Step 4:

In a dry vessel,2-(4-(6,7-dimethoxyquinazolin-4-yl)piperazin-1-yl)pyrimidin-5-ol (35 mg,0.095 mmol) (see method F), 1-(bromomethyl)-4-(trifluoromethyl)benzene(25 mg, 0.105 mmol), and potassium carbonate (20 mg, 0.142 mmol) weredissolved in dimethylformamide (1 mL). The reaction mixture was stirredat room temperature for 15 hours. The mixture was then diluted withwater and the resulting solids were filtered, washed with water anddried. The residue was purified by silica gel chromatography (0-6%methanol/methylene chloride. After drying on high vacuum, the product6,7-dimethoxy-4-(4-(5-((4-(trifluoromethyl)benzyl)oxy)pyrimidin-2-yl)piperazin-1-yl)quinazolinewas obtained as a white solid (18 mg, 36% yield).

Example 5

In a sealable reaction tube,4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline (75mg, 0.174 mmol), benzyl amine (0.19 mL, 1.74 mmol), copper iodide (3.5mg, 0.017 mmol), L-proline (4.0 mg, 0.035 mmol), and potassium phosphate(74 mg, 0.35 mmol), were combined in anhydrous dimethylsulfoxide (0.87mL) in a N₂ atmosphere and heated to 80 degrees Celsius for 15 hours.The cooled reaction mixture was diluted with ethyl acetate and filteredthrough a celite plug. The residue dissolved in ethylacetate was washedmultiple times with water. The residue was purified by silica gelchromatography (0-6% methanol/methylene chloride. After drying on highvacuum, the productN-benzyl-2-(4-(6,7-dimethoxyquinazolin-4-yl)piperazin-1-yl)pyrimidin-5-aminewas obtained as an off-white solid (38 mg, 48% yield).

Example 6

In a sealable reaction tube,4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline (50mg, 0.116 mmol), aniline (0.016 mL, 0.174 mmol), potassium carbonate (25mg, 0.174 mmol),chloro{[BrettPhos][2-(2-aminoethylphenyl]palladium(II)]}/[BrettPhos]admixture(molar PdP/P=1:1) (3.5 mg, 0.002 mmol), were combined in tert-butanol(0.8 mL) in a N₂ atmosphere and heated to 100 degrees Celsius for 15hours. The cooled reaction mixture was diluted with ethyl acetate andfiltered through a celite plug. The residue was purified by silica gelchromatography (0-6% methanol/methylene chloride. After drying on highvacuum, the product2-(4-(6,7-dimethoxyquinazolin-4-yl)piperazin-1-yl)-N-phenylpyrimidin-5-aminewas obtained as an off-white solid (45 mg, 88% yield).

Example 7

To a sealable vessel was added4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline (43mg, 0.1 mmol), Potassium (2-phenylethyl)trifluoroborate (25 mg, 0.12mmol), RuPhos (4.7 mg, 10 mol %), Pd(OAc)₂ (1.1 mg, 5 mol %), andpotassium carbonate (41 mg, 0.3 mmol). The reaction vessel was sealedand purged with nitrogen. 0.9 mL of toluene was added followed by 0.1 mLwater and the reaction vessel heated in an oilbath at 80° C. for 4hours. The reaction mixture was cooled and diluted with EtOAc and theorganic layer filtered through a plug of sodium sulfate. Evaporationgave the crude product which was subjected to flash chromatography usinga gradient of 40 to 100% EtOAc/Hex. Clean fractions were combined andevaporated to give 37.5 mg (82%) of the referenced compound as a whitesolid.

Example 8

In a microwave vessel,4-(4-(5-bromopyrimidin-2-yl)piperazin-1-yl)-6,7-dimethoxyquinazoline (50mg, 0.116 mmol), phenol (22 mg, 0.233 mmol), copper iodide (7.0 mg,0.035 mmol), 1,10-phenanthroline (11 mg, 0.06 mmol), and cesiumcarbonate (80 mg, 0.253 mmol), were combined in anhydrous dioxane (0.5mL) in a N₂ atmosphere and heated to 200 degrees Celsius for 1 hour in amicrowave reactor. The cooled reaction mixture was diluted with ethylacetate and filtered through a celite plug. The residue was purified bysilica gel chromatography (0-7% methanol/methylene chloride. Afterdrying on high vacuum, the product6,7-dimethoxy-4-(4-(5-phenoxypyrimidin-2-yl)piperazin-1-yl)quinazolinewas obtained as an off-white solid (13.5 mg, 26% yield).

Example 9

Step 1:

To a nitrogen-degassed mixture of (2-chloropyrimidin-5-yl)boronic acid(1.61 g, 10.2 mmol), bis(triphenylphosphine)palladium(II)dichloride(0.36 g, 0.51 mmol), sodium carbonate (3.24 g, 30.6 mmol), water (6.5mL), and dioxane (16 mL) was added benzyl bromide (1.92 g, 11.2 mmol).The mixture was stirred about 22 hours at 100° C. The reaction mixturewas cooled to room temperature, diluted with ethyl acetate (80 mL), andwashed with water (60 mL) and then brine (50 mL), then dried over sodiumsulfate, filtered, and evaporated under reduced pressure. The crudeamber oil was purified by silica gel chromatography with hexanes to 30%ethyl acetate/hexanes gradient to yield the product5-benzyl-2-chloropyrimidine as an orange oil which solidified uponstanding (1.21 g, 58% yield).

Step 2:

A mixture of 5-benzyl-2-chloropyrimidine (76 mg, 0.37 mmol),(1R,5S)-tert-butyl 3,8-diazabicyclo[3.2.1]octane-3-carboxylate (94 mg,0.44 mmol), and triethylamine (149 mg, 1.48 mmol) in 2-propanol (1 mL)was stirred in a sealed tube at 120° for 16 hours. The reaction mixturewas cooled to room temperature and the solvents were removed underreduced pressure. The crude residue was treated with water (6 mL), thensonicated and stirred. A light beige solid was isolated by filtration toyield the desired product (1R,5S)-tert-butyl8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate(111 mg, 79% yield).

Step 3:

A mixture of (1R,5S)-tert-butyl8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane-3-carboxylate(111 mg, 0.291 mmol) in 4N hydrochloric acid/dioxane (1.3 mL) wasstirred 2 hours at room temperature. The solvent was removed underreduced pressure. The amber solid was dried under high vacuum to yieldthe desired product(1R,5S)-8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane,assumed to be the di-HCl salt. Since the tare on the vial was lost, theyield was assumed to be quantitative (103 mg).

To a stirred 0° C. solution of tert-butyl2-(hydroxymethyl)morpholine-4-carboxylate (54 mg, 0.25 mmol) andtriethylamine (33 mg, 0.33 mmol) in dichloromethane (2 mL) was addedmethanesulfonyl chloride (29 mg, 0.25 mmol). The reaction mixture wasallowed to warm to room temperature and was stirred overnight for atotal of 22 hours. The mixture was diluted with dichloromethane (3 mL)and washed with aqueous 2N sodium hydroxide solution (3 mL). The organiclayers was dried over sodium sulfate, filtered, concentrated down, anddried under vacuum to yield the desired product tert-butyl2-(((methylsulfonyl)oxy)methyl)morpholine-4-carboxylate in excess yield.The yield was assumed to be quantitative (74 mg) and was used as is inthe next Example.

Example 10

Step 1:

A mixture of(1R,5S)-8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane di-HClsalt (70 mg, 0.2 mmol), commercially available4-chloro-6-hydroxy-7-methoxyquinazoline (38 mg, 0.18 mmol) andtriethylamine (109 mg, 1.08 mmol) in 2-propanol (1.5 mL) was stirred ina sealed tube at 120° C. for 1.5 hours. The reaction mixture was thencooled to room temperature. An off-white solid was isolated byfiltration and dried to yield desired product4-((1R,5S)-8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-methoxyquinazolin-6-olin slight excess yield (theoretical yield: 82 mg). The crude productprobably contained some triethylamine hydrochloride salt and was used asis for the next step. LCMS (M+1)=455.2.

Step 2:

A mixture of4-((1R,5S)-8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-methoxyquinazolin-6-ol(46 mg, <0.10 mmol), tert-butyl2-(((methylsulfonyl)oxy)methyl)morpholine-4-carboxylate (˜30 mg, 0.10mmol), and cesium carbonate (131 mg, 0.40 mmol) in N,N-dimethylformamide(0.3 mL), was stirred in a sealed tube at 85° C. for 4 hours. Thereaction mixture was treated with water (˜7 mL). A beige solid wasisolated by filtration, dried, and purified by silica gel preparativeTLC (elution with 10% methanol/dichloromethane) to yield the desiredproduct tert-butyl2-(((4-((1R,5S)-8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-methoxyquinazolin-6-yl)oxy)methyl)morpholine-4-carboxylateas a clear film (44.2 mg, 67%).

Example 11

A mixture of 5-benzyl-2-(piperazin-1-yl)pyrimidine di-HCl salt (45 mg,1.39 mmol), commercially available4-chloro-6-hydroxy-7-methoxyquinazoline (266 mg, 1.26 mmol) andtriethylamine (511 mg, 5.05 mmol) in 2-propanol (4 mL) was stirred in asealed tube at 120° C. for 6 hours. The reaction mixture was then cooledto room temperature. A beige solid was isolated by filtration and driedto yield desired product4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-7-methoxyquinazolin-6-ol inexcess yield (683 mg versus theoretical yield of 541 mg). The crudeproduct probably contained some triethylamine hydrochloride salt and wasused as is for the next step.

Example 12

A mixture of(1R,5S)-8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octane di-HClsalt (31 mg, 0.09 mmol), commercially available4-chloro-6,7-dimethoxyquinazoline (18 mg, 0.08 mmol) and triethylamine(49 mg, 0.48 mmol) in 2-propanol (0.7 mL) was stirred in a sealed tubeat 120° C. for 16 hours. The reaction mixture was then cooled to roomtemperature. A light-beige solid was isolated by filtration, washed witha small amount of water, and dried to yield4-((1R,5S)-8-(5-Benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-6,7-dimethoxyquinazoline(26 mg, 69% yield).

Example 13

A solution of tert-butyl2-((((4-((1R,5S)-8-(5-benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-methoxyquinazolin-6-yl)oxy)methyl)morpholine-4-carboxylate(44.2 mg, 0.068 mmol) in trifluoroacetic acid (1 mL) and dichloromethane(2 mL) was stirred 40 hours at room temperature. The excess solventswere evaporated. The residue was dissolved into dichloromethane (3 mL)and washed with aqueous 2N sodium hydroxide (3 mL). The aqueous layerwas extracted with fresh dichloromethane (4×3 mL). The combined organiclayers were dried over sodium sulfate, filtered, concentrated down, anddried to yield2-(((4-((1R,5S)-8-(5-Benzylpyrimidin-2-yl)-3,8-diazabicyclo[3.2.1]octan-3-yl)-7-methoxyquinazolin-6-yl)oxy)methyl)morpholineas a cloudy film (33.6 mg, 90% yield).

Example 14

A mixture of4-(4-(5-benzylpyrimidin-2-yl)piperazin-1-yl)-7-methoxyquinazolin-6-ol(47 mg, 0.11 mmol), 3-(4-methylpiperazin-1-yl)propyl methanesulfonate(35 mg, 0.147 mmol), and cesium carbonate (48 mg, 0.15 mmol) inN,N-dimethylformamide (0.2 mL) was stirred in a sealed tube at 85° C.for 18 hours. The reaction mixture was cooled to room temperature,treated with water (5 mL), sonicated, and stirred. The supernatant wasremoved, and the sticky residue was dissolved into dichloromethane,dried over sodium sulfate, filtered, and concentrated down. The cruderesidue was purified by silica gel preparative TLC (elution system was88:12:1 dichloromethane/methanol/ammonium hydroxide) to yield4-(4-(5-Benzylpyrimidin-2-yl)piperazin-1-yl)-7-methoxy-6-(3-(4-methylpiperazin-1-yl)propoxy)quinazolineas a beige foamy solid (15.4 mg, 25% yield).

NMR and LC MS data for compounds disclosed herein are shown below.

LC Compound MS Number 1H NMR (M + 1) 1 1H NMR (400 MHz, DMSO-d6) δ 8.31(s, 2H), 8.23 (s, 1H), 8.13 (s, 1H), 372.4 7.34-7.13 (m, 5H), 4.44-4.10(m, 4H), 3.90-3.73 (m, 6H). 2 ¹H NMR (400 MHz, DMSO-d₆) δ 8.31 (s, 1H),8.30 (s, 1H), 7.40-7.09 (m, 5H), 373.5 4.03-3.97 (m, 2H), 3.89 (b.s.,1H), 3.88-3.82 (m, 2H), 3.81-3.72 (m, 6H), 3.72-3.65 (m, 4H). 3Reference compound; not encompassed within the current invention. 382.54 1H NMR (400 MHz, DMSO-d6) δ 8.66 (s, 1H), 8.32 (s, 2H), 8.11-7.99 (m,1H), 412.5 7.35-7.12 (m, 7H), 3.92 (broad singlet, 8H), 3.80 (s, 2H). 51H NMR (400 MHz, DMSO-d6) δ 8.78 (s, 1H), 8.34 (s, 2H), 7.85 (d, J = 9.2Hz, 412.5 1H), 7.67 (dd, J = 9.2, 2.6 Hz, 1H), 7.47 (d, J = 2.7 Hz, 1H),7.38-7.04 (m, 5H), 4.19 (m, 4H), 4.03-3.86 (m, 7H), 3.81 (s, 2H). 6 ¹HNMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.23 (s, 2H), 7.21 (d, J = 13.1Hz, 422.5 2H), 3.92 (s, 6H), 3.84 (m, 6H), 3.72-3.65 (m, 4H), 1.24-1.14(m, 1H), 0.59-0.51 (m, 2H), 0.33-0.26 (m, 2H). 7 1H NMR (400 MHz,DMSO-d6) δ 8.59 (s, 1H), 8.32 (s, 2H), 7.79 (d, J = 9.1 Hz, 426.5 1H),7.50 (dd, J = 9.1, 2.7 Hz, 1H), 7.34-7.15 (m, 6H), 4.89 (m 1H), 4.48 (m,1H), 4.26-4.09 (m, 2H), 3.92 (s, 3H), 3.79 (s, 2H), 3.46 (m, 1H), 3.37(dd, J = 13.1, 3.8 Hz, 1H), 3.26-3.13 (m, 1H), 1.29 (d, J = 6.6 Hz, 3H).8 1H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 8.29 (s, 2H), 7.97 (d, J =9.2 Hz, 428.5 1H), 7.47-7.29 (m, 5H), 7.20 (d, J = 2.7 Hz, 1H), 7.15(dd, J = 9.2, 2.7 Hz, 1H), 5.11 (s, 2H), 3.91 (s, 3H), 3.86-3.80 (m,5H), 3.75 (m, 4H). 9 ¹H NMR (400 MHz, Chloroform-d) δ 8.71 (s, 1H), 8.61(s, 2H), 7.53-7.43 (m, 428.5 4H), 7.39-7.29 (m, 2H), 7.18 (s, 1H),4.15-4.09 (m, 4H), 4.05 (s, 3H), 4.01 (s, 3H), 3.81 (m, 4H). 10 ¹H NMR(400 MHz, DMSO-d6) δ 8.59 (s, 1H), 8.29 (s, 2H), 7.78 (d, J = 9.1 Hz,428.5 1H), 7.50 (dd, J = 9.1, 2.8 Hz, 1H), 7.47-7.30 (m, 5H), 7.26 (d, J= 2.8 Hz, 1H), 5.12 (s, 2H), 3.91 (s, 3H), 3.89-3.82 (m, 4H), 3.78-3.71(m, 4H). 11 ¹H NMR (400 MHz, Chloroform-d) δ 8.70 (s, 1H), 8.23 (s, 2H),7.32-7.27 (m, 442.5 3H), 7.23-7.16 (m, 4H), 4.08-3.97 (m, 10H), 3.82 (s,2H), 3.74 (m, 4H). 12 1H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.27 (d,J = 4.9 Hz, 1H), 7.30 (m, 442.5 4H), 7.27-7.16 (m, 3H), 6.51 (d, J = 4.9Hz, 1H), 4.02-3.84 (m, 12H), 3.74-3.63 (m, 4H). 13 ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (s, 1H), 8.31 (s, 2H), 7.72 (s, 1H), 7.22 (m, 443.5 2H),7.17-7.11 (m, 2H), 6.81-6.76 (m, 2H), 6.72-6.66 (m, 1H), 3.97-3.89 (m,10H), 3.75-3.69 (m, 4H). 14 ¹H NMR (400 MHz, DMSO-d₆) δ 8.57 (s, 1H),8.35 (s, 2H), 7.38-7.31 (m, 2H), 444.5 7.23 (m, 2H), 7.08 (m, 1H),7.01-6.94 (m, 2H), 4.00-3.89 (m, 10H), 3.79-3.69 (m, 4H). 15 1H NMR (400MHz, DMSO-d6) δ 8.77 (s, 1H), 8.67 (s, 2H), 8.56 (s, 1H), 8.15 (s, 444.51H), 8.05 (dd, J = 5.3, 1.9 Hz, 1H), 7.52 (ddd, J = 8.7, 7.1, 2.0 Hz,1H), 7.22 (d, J = 9.0 Hz, 2H), 6.74-6.65 (m, 2H), 4.00-3.83 (m, 10H),3.71 (t, J = 5.2 Hz, 4H). 16 1H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H),8.35 (s, 2H), 8.14 (t, J = 1.8 Hz, 444.5 1H), 7.97-7.83 (m, 2H), 7.22(d, J = 9.0 Hz, 2H), 7.13 (m, 2H), 3.93 (br.s, 10H), 3.79-3.64 (m, 4H).17 444.5 18 ¹H NMR (400 MHz, DMSO-d₆) δ 8.56 (s, 1H), 8.24 (s, 2H), 7.21(d, J = 13.0 Hz, 450.5 2H), 3.93 (s, 6H), 3.85 (m, 6H), 3.72-3.64 (m,4H), 2.27 (m, 1H), 1.80-1.69 (m, 2H), 1.55 (m, 4H), 1.37-1.24 (m, 2H).19 ¹H NMR (400 MHz, Chloroform-d) δ 8.70 (s, 1H), 8.62 (s, 2H),7.77-7.72 (m, 453.5 2H), 7.63-7.58 (m, 3H), 7.17 (s, 1H), 4.17-4.11 (m,4H), 4.06 (s, 3H), 4.01 (s, 3H), 3.87 (m, 4H). 20 455.5 21 ¹H NMR (400MHz, DMSO-d₆) δ 8.73 (s, 2H), 8.57 (s, 1H), 7.38-7.17 (m, 7H), 456.54.42 (s, 2H), 4.13-4.03 (m, 4H), 3.99 (s, 2H), 3.93 (s, 3H), 3.92 (s,3H). 22 ¹H NMR (400 MHz, DMSO-d₆) δ 8.56 (s, 1H), 8.29 (s, 2H), 7.21 (m,2H), 7.10 (m, 456.5 4H), 3.93 (s, 3H), 3.92 (s, 3H), 3.90 (m, 4H), 3.74(s, 2H), 3.72-3.67 (m, 4H), 2.24 (s, 3H). 23 1H NMR (400 MHz, DMSO-d6) δ8.56 (s, 1H), 8.31 (s, 2H), 7.33-7.14 (m, 7H), 456.5 4.88 (m, 1H), 4.49(m, 1H), 4.19-4.03 (m, 2H), 3.93, (s, 6H), 3.79 (s, 2H), 3.43 (td, J =13.8, 12.7, 3.3 Hz, 1H), 3.14 (td, J = 12.4, 3.5 Hz, 1H), 1.31 (d, J =6.6 Hz, 3H). 24 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.22 (s, 2H),7.32-7.11 (m, 7H), 456.5 3.97-3.84 (m, 10H), 3.74-3.63 (m, 4H),2.87-2.79 (m, 2H), 2.77-2.69 (m, 2H). 25 ¹H NMR (400 MHz, DMSO-d₆) δ8.56 (s, 1H), 8.23 (s, 2H), 7.21 (m, 2H), 456.5 7.18-7.08 (m, 4H), 3.93(s, 3H), 3.92 (s, 3H), 3.90 (m, 4H), 3.79 (s, 2H), 3.73-3.67 (m, 4H),2.24 (s, 3H). 26 1H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.31 (s, 2H),7.34-7.14 (m, 7H), 456.5 4.88 (m, 1H), 4.49 (m, 1H), 4.20-4.02 (m, 2H),3.93 (s, 6H), 3.79 (s, 2H), 3.43 (m, 1H), 3.20-3.08 (m, 1H), 1.31 (d, J= 6.6 Hz, 3H). 27 ¹H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.29 (s,2H), 7.35-7.10 (m, 7H), 456.5 4.60-4.47 (m, 2H), 4.40-4.31 (m, 1H), 3.92(m, 7H), 3.78 (s, 2H), 3.60-3.48 (m, 1H), 3.40 (dd, J = 13.2, 3.5 Hz,1H), 1.22 (d, J = 6.3 Hz, 3H). 1 proton partially buried under waterpeak. 28 ¹H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 8.29 (s, 2H),7.33-7.11 (m, 7H), 456.5 4.59-4.46 (m, 2H), 4.36 (dt, J = 13.1, 1.9 Hz,1H), 3.92 (d, J = 2.3 Hz, 7H), 3.78 (s, 2H), 3.53 (ddd, J = 13.7, 11.3,3.3 Hz, 1H), 3.40 (dd, J = 13.1, 3.6 Hz, 1H), 1.22 (d, J = 6.5 Hz, 3H)—1proton partially buried under water peak. 29 ¹H NMR (400 MHz, DMSO-d₆) δ8.54 (s, 1H), 7.65 (s, 1H), 7.32-7.13 (m, 7H), 457.5 6.30 (b.s., 2H),3.92 (s, 3H), 3.91 (s, 3H), 3.86-3.81 (m, 4H), 3.67 3.60 (m, 6H). 30 1HNMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 8.31 (s, 2H), 7.69 (s, 1H), 7.25(d, 457.5 J = 5.4 Hz, 2H), 7.18-7.09 (m, 2H), 6.81-6.73 (m, 2H), 6.69(m, 1H), 4.88 (m, 1H), 4.54-4.41 (m, 1H), 4.21-4.06 (m, 3H), 3.94 (2closely spaced singlets, total of 6H), 3.52-3.39 (m, 1H), 3.18 (td, J =12.2, 3.5 Hz, 1H), 1.34 (d, J = 6.5 Hz, 3H). 31 ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (s, 1H), 8.28 (s, 2H), 7.58 (s, 1H), 7.22 (d, J = 10.1Hz, 457.5 2H), 6.97 (d, J = 8.3 Hz, 2H), 6.77-6.68 (m, 2H), 3.93 (s,6H), 3.90 (m, 4H), 3.74-3.68 (m, 4H), 2.17 (s, 3H). 32 ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (s, 1H), 8.26 (s, 2H), 7.22 (d, J = 8.9 Hz, 457.5 2H),7.08 (d, J = 7.6 Hz, 1H), 7.01-6.95 (m, 1H), 6.92 (s, 1H), 6.74-6.65 (m,2H), 3.92 (m, 10H), 3.72 (m, 4H), 2.21 (s, 3H). 33 ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (s, 1H), 8.30 (s, 2H), 7.64 (s, 1H), 7.22 (d, J = 9.1Hz, 457.5 2H), 7.02 (t, J = 7.6 Hz, 1H), 6.58 (d, J = 8.7 Hz, 2H), 6.52(d, J = 7.4 Hz, 1H), 3.99-3.87 (m, 10H), 3.77-3.67 (m, 4H), 2.19 (s,3H). 34 ¹H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 7.93 (s, 2H),7.39-7.28 (m, 4H), 457.5 7.26-7.15 (m, 3H), 5.86 (t, J = 6.2 Hz, 1H),4.23 (d, J = 6.3 Hz, 2H), 3.92 (s, 3H), 3.91 (s, 3H), 3.73 (m, 4H), 3.64(m, 4H). 35 1H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 7.63 (s, 1H),7.32-7.07 (m, 7H), 458.5 3.92 (2 closely spaced singlets, 6H), 3.78(br.s, 4H), 3.69 (m, 4H), 3.57 (s, 2H). 36 1H NMR (400 MHz, DMSO-d6) δ8.51 (s, 2H), 7.29 (dd, J = 8.7, 7.2 Hz, 2H), 458.5 7.22 (d, J = 8.8 Hz,2H), 7.05-6.98 (m, 2H), 6.98-6.89 (m, 1H), 4.95 (s, 2H), 3.98 (m, 4H),3.93 (s, 6H), 3.72 (m, 4H). 37 ¹H NMR (400 MHz, Chloroform-d) δ 8.69 (s,1H), 8.17 (s, 2H), 7.45-7.31 (m, 458.5 6H), 7.16 (s, 1H), 5.05 (s, 2H),4.04 (s, 3H), 3.99 (s, 3H), 4.00-3.92 (m, 4H), 3.76 (m, 4H). 38 ¹H NMR(400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.31 (s, 2H), 7.30-7.24 (m, 2H),460.5 7.21 (m, 2H), 7.14-7.07 (m, 2H), 3.91 (m, 10H), 3.79 (s, 2H),3.74-3.63 (m, 4H). 39 ¹H NMR (400 MHz, Chloroform-d) δ 8.66 (s, 1H),8.22 (s, 2H), 7.25-7.15 (m, 460.5 4H), 7.00-6.81 (m, 2H), 4.09-3.93 (s,14H), 3.82 (s, 2H). 40 ¹H NMR (400 MHz, DMSO-d₆) δ 8.58 (s, 1H), 8.53(s, 2H), 7.30 (m, 2H), 460.5 7.27-7.11 (m, 5H), 4.02 (m, 4H), 3.93 (s,6H), 3.75 (m, 4H). 41 1H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.28 (s,1H), 7.22 (d, J = 12.1 Hz, 461.3 2H), 3.94 (m, 13H), 3.70 (m, 4H). 42 ¹HNMR (400 MHz, DMSO-d₆) δ 8.56 (s, 1H), 8.29 (s, 2H), 7.69 (s, 1H), 7.22(d, J = 10.4 Hz, 461.5 2H), 7.03-6.94 (m, 2H), 6.84-6.75 (m, 2H),3.95-3.88 (m, 10H), 3.74-3.68 (m, 4H). 43 ¹H NMR (400 MHz, DMSO-d₆) δ8.56 (s, 1H), 8.28 (s, 2H), 7.56 (dd, J = 5.1, 1.3 Hz, 464.5 1H),7.29-7.14 (m, 2H), 7.02 (dd, J = 5.1, 3.4 Hz, 1H), 5.30 (s, 2H), 3.93(s, 3H), 3.92 (s, 3H), 3.86 (dd, J = 7.2, 3.4 Hz, 4H), 3.68 (dd, J =6.2, 4.1 Hz, 4H). 44 ¹H NMR (400 MHz, DMSO-d₆) δ 8.57 (s, 1H), 8.27 (s,2H), 7.56 (m, 1H), 464.5 7.30-7.10 (m, 3H), 5.10 (s, 2H), 3.93 (s, 3H),3.92 (s, 3H), 3.85 (m, 4H), 3.72-3.65 (m, 4H). 45 ¹H NMR (400 MHz,DMSO-d₆) δ 8.55 (s, 1H), 8.34 (s, 2H), 7.76 (d, J = 7.9 Hz, 467.5 2H),7.45 (d, J = 7.9 Hz, 2H), 7.21 (d, J = 13.1 Hz, 2H), 4.00-3.84 (m, 12H),3.69 (dd, J = 6.9, 3.5 Hz, 4H). 46 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s,1H), 8.35 (s, 2H), 7.75 (t, J = 1.7 Hz, 467.5 1H), 7.67 (dt, J = 7.7,1.4 Hz, 1H), 7.60 (dt, J = 7.9, 1.5 Hz, 1H), 7.50 (t, J = 7.7 Hz, 1H),7.21 (d, J = 13.2 Hz, 2H), 3.96-3.88 (m, 10H), 3.86 (s, 2H), 3.72-3.65(m, 4H). 47 ¹H NMR (400 MHz, DMSO-d₆) δ 8.57 (s, 1H), 8.51 (s, 1H), 8.35(s, 2H), 7.52 (d, J = 8.8 Hz, 468.5 2H), 7.22 (d, J = 9.1 Hz, 2H), 6.79(d, J = 8.8 Hz, 2H), 4.00-3.89 (m, 10H), 3.76-3.69 (m, 4H). 48 ¹H NMR(400 MHz, DMSO-d₆) δ 8.45 (s, 1H), 8.34 (s, 2H), 7.34-7.11 (m, 7H),468.6 4.76 (s, 2H), 4.21 (d, J = 12.5 Hz, 2H), 3.90 (s, 3H), 3.88 (s,3H), 3.80 (s, 2H), 3.48 (d, J = 12.6 Hz, 2H), 3.18-3.00 (m, 2H), 1.16(t, J = 7.3 Hz, 2H). 49 1H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 8.47(s, 2H), 7.23 (d, J = 8.2 Hz, 469.5 3H), 7.15 (d, J = 7.4 Hz, 1H), 6.99(t, J = 7.7 Hz, 1H), 6.74-6.59 (m, 2H), 3.94 (m, 10H), 3.84 (t, J = 8.5Hz, 2H), 3.72 (m, 4H), 3.07 (t, J = 8.3 Hz, 2H). 50 470.6 51 1H NMR (400MHz, DMSO-d6) δ 10.10 (s, 1H), 8.94 (s, 2H), 8.57 (s, 1H), 471.57.77-7.67 (m, 2H), 7.34 (t, J = 7.9 Hz, 2H), 7.23 (d, J = 6.6 Hz, 2H),7.09 (t, J = 7.4 Hz, 1H), 4.18-4.02 (m, 4H), 3.94 (two closely spacedsinglets, 3H each), 3.85-3.65 (m, 4H). 52 ¹H NMR (400 MHz,DMSO-d_(6) δ 8.64 (s, 1H), 8.11 (s, 2H), 7.35-7.28 (m, 3H),) 471.57.27-7.19 (m, 4H), 4.42 (s, 2H), 3.93 (s, 3H), 3.92 (s, 3H), 3.79 (dd, J= 6.8, 3.5 Hz, 4H), 3.69-3.64 (m, 4H), 2.88 (s, 3H). 53 ¹H NMR (400 MHz,DMSO-d6) δ 8.55 (s, 1H), 7.91 (s, 2H), 7.38-7.00 (m, 6H), 471.5 5.81 (t,J = 6.4 Hz, 1H), 4.18 (d, J = 6.2 Hz, 2H), 3.92 (s, 3H), 3.91 (s, 3H),3.78 (m, 4H), 3.65 (m, 4H), 2.26 (s, 3H). 54 ¹H NMR (400 MHz, DMSO-d6) δ8.55 (s, 1H), 7.95 (s, 2H), 7.31-7.10 (m, 6H), 471.6 5.67 (t, J = 5.9Hz, 1H), 4.18 (d, J = 5.7 Hz, 2H), 3.92 (s, 3H), 3.91 (s, 3H), 3.75 (m,4H), 3.65 (m, 4H), 2.31 (s, 3H). 55 ¹H NMR (400 MHz, DMSO-d6) δ 8.56 (s,1H), 7.92 (s, 2H), 7.36-6.95 (m, 6H), 471.6 5.82 (t, J = 6.2 Hz, 1H),4.19 (d, J = 6.2 Hz, 2H), 3.92 (s, 3H), 3.91 (s, 3H), 3.73 (m, 4H), 3.65(m, 4H), 2.28 (s, 3H). 56 ¹H NMR (400 MHz, DMSO-d6) δ 8.58 (s, 1H), 7.82(s, 2H), 7.46-7.05 (m, 7H), 471.6 5.82 (d, J = 7.5 Hz, 1H), 4.43 (p, J =6.8 Hz, 1H), 3.92 (s, 3H), 3.91 (s, 3H), 3.69 (m, 4H), 3.63 (m, 4H),1.40 (d, J = 6.6 Hz, 3H). 57 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H),8.32 (s, 2H), 7.25-7.16 (m, 3H), 472.5 6.84-6.72 (m, 3H), 3.92 (m, 10H),3.75 (s, 2H), 3.73-3.64 (m, 7H). 58 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s,1H), 8.29 (s, 2H), 7.21 (d, J = 12.7 Hz, 472.5 2H), 7.16-7.11 (m, 2H),6.87-6.82 (m, 2H), 3.97-3.87 (m, 10H), 3.72 (s, 2H), 3.69 (m, 7H). 59 ¹HNMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.15 (s, 2H), 7.42-7.32 (m, 4H),472.5 7.29-7.24 (m, 1H), 7.22-7.17 (m, 2H), 5.42 (q, J = 6.3 Hz, 1H),3.92 (s, 3H), 3.90 (s, 3H), 3.80 (m, 4H), 3.68-3.61 (m, 4H), 1.55 (d, J= 6.3 Hz, 3H). 60 1H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.29 (s,2H), 7.48-7.28 (m, 5H), 472.5 7.24 (br.s, 2H), 5.11 (s, 2H), 4.81 (d, J= 6.1 Hz, 1H), 4.39 (dd, J = 13.2, 3.4 Hz, 1H), 4.18-4.04 (m, 2H), 3.93(s, 6H), 3.47-3.35 (m, 1H), 3.14 (td, J = 12.1, 3.3 Hz, 1H), 1.29 (d, J= 6.6 Hz, 3H). 61 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.28 (s,2H), 7.33-7.10 (m, 6H), 472.5 5.07 (s, 2H), 3.92 (s, 6H), 3.85 (dd, J =7.0, 3.6 Hz, 4H), 3.75-3.63 (m, 4H), 2.31 (s, 3H). 62 ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (s, 1H), 8.27 (s, 2H), 7.35-7.27 (m, 2H), 472.57.25-7.14 (m, 4H), 5.06 (s, 2H), 3.92 (s, 6H), 3.88-3.80 (m, 4H),3.73-3.64 (m, 4H), 2.29 (s, 3H). 63 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s,1H), 8.15 (s, 2H), 7.42-7.31 (m, 4H), 472.5 7.29-7.15 (m, 3H), 5.42 (q,J = 6.4 Hz, 1H), 3.92 (s, 3H), 3.90 (s, 3H), 3.80 (dd, J = 6.6, 3.7 Hz,4H), 3.69-3.61 (m, 4H), 1.55 (d, J = 6.4 Hz, 3H). 64 ¹H NMR (400 MHz,DMSO-d₆) δ 8.54 (s, 1H), 8.15 (s, 2H), 7.41-7.38 (m, 2H), 472.5 7.34 (m,2H), 7.30-7.24 (m, 1H), 7.22-7.17 (m, 2H), 5.42 (q, J = 6.4 Hz, 1H),3.92 (s, 3H), 3.90 (s, 3H), 3.80 (m, 4H), 3.68-3.62 (m, 4H), 1.55 (d, J= 6.4 Hz, 3H). 65 ¹H NMR (400 MHz, DMSO-d₆) δ 8.56 (s, 1H), 8.31 (s,2H), 7.40-7.37 (m, 1H), 472.5 7.27-7.16 (m, 5H), 5.75 (s, 1H), 5.11 (s,2H), 3.93 (s, 6H), 3.89-3.83 (m, 4H), 3.74-3.67 (m, 4H), 2.33 (s, 3H).66 1H NMR (400 MHz, DMSO-d6) δ 8.56 (s, 1H), 8.29 (s, 2H), 7.48-7.28 (m,5H), 472.5 7.24 (br.s, 2H), 5.11 (s, 2H), 4.81 (d, J = 7.6 Hz, 1H), 4.39(d, J = 13.2 Hz, 1H), 4.10 (dd, J = 15.8, 12.8 Hz, 2H), 3.93 (s, 6H),3.48-3.35 (m, 1H), 3.14 (td, J = 12.2, 3.6 Hz, 1H), 1.29 (d, J = 6.6 Hz,3H). 67 ¹H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.26 (s, 1H), 7.39 (m,5H), 7.22 (s, 472.5 1H), 7.15 (s, 1H), 5.10 (s, 2H), 4.53 (d, J = 6.7Hz, 1H), 4.43 (d, J = 12.9 Hz, 1H), 4.33-4.21 (m, 1H), 3.92 (d, J = 2.1Hz, 7H), 3.54 (t, J = 11.4 Hz, 1H), 3.38 (dd, J = 13.2, 3.6 Hz, 1H),3.31-3.19 (m, 1H), 1.23 (d, J = 6.6 Hz, 3H). 68 1H NMR (400 MHz,DMSO-d6) δ 8.55 (s, 1H), 8.26 (s, 2H), 7.48-7.29 (m, 5H), 472.5 7.22 (s,1H), 7.15 (s, 1H), 5.10 (s, 2H), 4.59-4.48 (m, 1H), 4.43 (d, J = 12.8Hz, 1H), 4.32-4.20 (m, 1H), 3.92 (s, 6H), 3.54 (td, J = 13.3, 12.5, 3.3Hz, 1H), 3.38 (dd, J = 13.0, 3.6 Hz, 1H), 3.24 (m, 1H), 1.23 (d, J = 6.6Hz, 3H). 69 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.29 (s, 2H),7.52-7.46 (m, 2H), 476.5 7.26-7.18 (m, 4H), 5.09 (s, 2H), 3.93 (s, 3H),3.92 (s, 3H), 3.88-3.83 (m, 4H), 3.71-3.66 (m, 4H). 70 ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (s, 1H), 8.31 (s, 2H), 7.87 (s, 1H), 7.22 (d, J = 9.3Hz, 478.0 2H), 7.19-7.12 (m, 2H), 6.82-6.71 (m, 2H), 3.93 (s, 10H),3.76-3.69 (m, 4H). 71 1H NMR (400 MHz, DMSO-d6) δ 8.55 (s, 1H), 8.33 (s,2H), 7.39-7.29 (m, 2H), 478.5 7.21 (d, J = 12.9 Hz, 2H), 7.12-7.04 (m,1H), 3.92 (m, 10H), 3.79 (s, 2H), 3.69 (m, 4H). 72 478.5 73 ¹H NMR (400MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.31 (s, 2H), 7.87 (d, J = 7.9 Hz, 483.52H), 7.64 (d, J = 8.0 Hz, 2H), 7.21 (d, J = 14.1 Hz, 2H), 5.23 (s, 2H),3.93 (s, 6H), 3.85 (m, 4H), 3.68 (t, J = 5.0 Hz, 4H). 74 1H NMR (400MHz, DMSO-d6) δ 8.57 (s, 1H), 8.36 (s, 2H), 7.23 (d, J = 7.6 Hz, 483.62H), 6.97 (d, J = 7.3 Hz, 1H), 6.85 (t, J = 7.9 Hz, 1H), 6.58 (dd, J =7.8, 6.6 Hz, 1H), 6.26 (d, J = 8.2 Hz, 1H), 4.02-3.87 (m, 10H), 3.73 (m,4H), 3.53-3.46 (m, 2H), 2.79 (t, J = 6.4 Hz, 2H), 2.02-1.92 (m, 2H). 75¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.34 (s, 2H), 7.94 (s, 1H),7.72 (m, 485.5 1H), 7.69 (dt, J = 7.2, 1.7 Hz, 1H), 7.42-7.37 (m, 1H),7.37-7.30 (m, 3H), 7.21 (d, J = 12.3 Hz, 2H), 3.92 (m, 10H), 3.84 (s,2H), 3.72-3.65 (m, 4H). 76 1H NMR (400 MHz, DMSO-d6) δ 10.11 (s, 1H),8.58 (s, 2H), 8.55 (s, 1H), 485.5 7.32 (m, 4H), 7.28-7.15 (m, 3H),4.00-3.85 (m, 10H), 3.74-3.65 (m, 4H), 3.62 (s, 2H). 77 ¹H NMR (400 MHz,DMSO-d₆) δ 8.56 (s, 1H), 8.26 (s, 2H), 7.40-7.32 (m, 2H), 488.5 7.21 (d,J = 13.5 Hz, 2H), 6.97-6.89 (m, 2H), 5.03 (s, 2H), 3.92 (s, 6H), 3.85(dd, J = 6.5, 3.8 Hz, 4H), 3.74 (s, 3H), 3.69 (dd, J = 6.5, 3.6 Hz, 4H).78 492.6 79 ¹H NMR (400 MHz, DMSO-d₆) δ 8.61-8.51 (m, 1H), 8.33-8.29 (m,2H), 497.6 7.37-7.11 (m, 7H), 4.24 (dd, J = 21.6, 6.7 Hz, 2H), 3.97-3.86(m, 7H), 3.78 (d, J = 10.2 Hz, 2H), 3.75-3.61 (m, 4H), 3.52-3.46 (m,1H), 3.17-2.98 (m, 4H), 2.90-2.80 (m, 1H). 80 ¹H NMR (400 MHz, DMSO-d₆)δ 8.55 (s, 1H), 8.32 (s, 2H), 7.39-7.11 (m, 7H), 497.6 5.00 (b.s., 1H),3.96-3.86 (m, 7H), 3.80 (s, 2H), 3.72-3.60 (m, 4H), 3.18-3.10 (m, 1H),3.03-2.89 (m, 2H), 2.90-2.75 (m, 1H), 2.15-2.02 (m, 1H), 1.94-1.82 (m,2H). 81 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.31 (s, 2H), 7.29 (d,J = 8.0 Hz, 498.6 2H), 7.21 (d, J = 12.6 Hz, 2H), 7.14 (d, J = 8.0 Hz,2H), 3.92 (m, 10H), 3.75 (s, 2H), 3.68 (dd, J = 6.7, 3.6 Hz, 4H), 1.23(s, 9H). 82 ¹H NMR (400 MHz, DMSO-d₆) δ 8.56 (s, 1H), 8.29 (s, 2H), 7.61(s, 1H), 7.22 (d, J = 9.5 Hz, 499.6 2H), 7.20-7.15 (m, 2H), 6.77-6.72(m, 2H), 3.95-3.87 (m, 10H), 3.74-3.68 (m, 4H), 1.22 (s, 9H). 83 ¹H NMR(400 MHz, DMSO-d6) δ 8.54 (s, 1H), 7.92 (s, 2H), 7.20 (m, 2H), 6.92 (s,501.5 1H), 6.84 (m, 2H), 5.96 (s, 2H), 5.79 (t, J = 6.3 Hz, 1H), 4.14(d, J = 6.3 Hz, 2H), 3.92 (s, 3H), 3.91 (s, 3H, 3.73 (m, 4H), 3.65 (m,4H). 84 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.31 (s, 2H),7.35-7.15 (m, 7H), 502.6 5.03 (d, J = 5.1 Hz, 1H), 4.72 (t, J = 5.6 Hz,1H), 4.17 (dd, J = 10.3, 4.0 Hz, 1H), 4.03 (dd, J = 10.3, 6.1 Hz, 1H),3.93 (s, 3H), 3.91 (dd, J = 6.6, 3.6 Hz, 4H), 3.86 (q, J = 5.2 Hz, 1H),3.80 (s, 2H), 3.70-3.63 (m, 4H), 3.46 (t, J = 5.8 Hz, 2H). 85 ¹H NMR(400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.32 (s, 2H), 7.37-7.14 (m, 7H),502.6 5.06 (s, 1H), 4.74 (s, 1H), 4.16 (dd, J = 10.2, 4.1 Hz, 1H), 4.03(dd, J = 10.2, 6.1 Hz, 1H), 3.94-3.92 (m, 7H), 3.90-3.83 (m, 1H), 3.80(s, 2H), 3.75-3.66 (m, 4H), 3.49-3.44 (m, 2H). 86 ¹H NMR (400 MHz,DMSO-d6) δ 8.55 (s, 1H), 8.32 (s, 2H), 7.35-7.06 (m, 6H), 508.5 3.92 (m,10H), 3.79 (s, 2H), 3.68 (m, 4H). 87 ¹H NMR (400 MHz, DMSO-d₆) δ 8.57(s, 1H), 8.32 (s, 2H), 7.77 (d, J = 9.1 Hz, 509.6 1H), 7.49 (dd, J =9.1, 2.7 Hz, 1H), 7.34-7.15 (m, 6H), 3.98 (d, J = 5.8 Hz, 2H), 3.94-3.88(m, 4H), 3.80 (s, 2H), 3.77-3.70 (m, 4H), 2.82-2.75 (m, 2H), 2.15 (s,3H), 1.94-1.81 (m, 1H), 1.80-1.67 (m, 4H), 1.41-1.28 (m, 2H). 88 ¹H NMR(400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.35 (s, 2H), 7.65 (d, J = 7.9 Hz,510.5 2H), 7.47 (d, J = 7.9 Hz, 2H), 7.21 (d, J = 12.6 Hz, 2H),3.97-3.87 (m, 12H), 3.69 (dd, J = 6.4, 3.6 Hz, 4H). 89 ¹H NMR (400 MHz,DMSO-d₆) δ 8.54 (s, 1H), 8.32 (s, 2H), 7.36-7.12 (m, 7H), 511.64.05-3.95 (m, 2H), 3.93 (s, 3H), 3.92-3.88 (m, 4H), 3.80 (s, 2H),3.72-3.62 (m, 4H), 2.94-2.78 (m, 2H), 2.75-2.62 (m, 3H), 1.90-1.78 (m,1H), 1.52-1.37 (m, 1H). 90 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H),8.32 (s, 2H), 7.33-7.15 (m, 7H), 511.6 4.26-4.21 (m, 2H), 3.99-3.88 (m,7H), 3.80 (s, 2H), 3.74-3.63 (m, 4H), 3.39-3.32 (m, 2H), 3.09-3.02 (m,2H), 2.87-2.78 (m, 1H), 2.25 (s, 3H). 91 511.6 92 ¹H NMR (400 MHz,DMSO-d₆) δ 8.55 (s, 1H), 8.32 (s, 1H), 7.38-7.10 (m, 7H), 511.63.96-3.85 (m, 7H), 3.80 (s, 2H), 3.72-3.63 (m, 4H), 3.45-3.35 (m, 2H),2.78-2.73 (m, 2H), 2.37-2.32 (m, 1H), 2.27 (s, 3H), 1.94-1.82, (m, 2H).93 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.31 (s, 2H), 7.36-7.12 (m,7H), 512.6 4.52 (d, J = 5.8 Hz, 2H), 4.33 (d, J = 5.8 Hz, 2H), 4.23 (s,2H), 3.92 (s, 3H), 3.92-3.87 (m, 4H), 3.79 (s, 2H), 3.74-3.62 (m, 4H),1.39 (s, 3H). 94 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.28 (s, 2H),7.43-7.33 (m, 4H), 514.6 7.21 (d, J = 14.3 Hz, 2H), 5.07 (s, 3H), 3.92(s, 6H), 3.85 (dd, J = 6.5, 3.7 Hz, 4H), 3.68 (dd, J = 6.3, 3.9 Hz, 4H),1.27 (s, 9H). 95 ¹H NMR (400 MHz, DMSO-d₆) δ 8.57 (s, 1H), 7.76 (d, J =9.1 Hz, 1H), 7.66 (s, 524.7 1H), 7.48 (dd, J = 9.1, 2.7 Hz, 1H),7.33-7.12 (m, 6H), 6.31 (s, 2H), 3.98 (d, J = 5.7 Hz, 2H), 3.87-3.80 (m,4H), 3.73-3.66 (m, 4H), 3.65 (s, 2H), 2.85-2.75 (m, 2H), 2.16 (s, 3H),1.95-1.82 (m, 2H), 1.81-1.69 (m, 3H), 1.43-1.27 (m, 2H). 96 525.6 97 ¹HNMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.32 (s, 2H), 7.35-7.14 (m, 7H),525.6 3.97 (d, J = 6.6 Hz, 2H), 3.93 (s, 3H), 3.92-3.87 (m, 4H), 3.80(s, 2H), 3.73-3.64 (m, 4H), 3.16-3.02 (m, 1H), 2.92-2.84 (m, 1H),2.47-2.34 (m, 1H), 2.07-1.90 (m, 1H), 1.88-1.78 (m, 1H), 1.66-1.56 (m,1H), 1.47-1.34 (m, 1H), 1.32-1.16 (m, 2H). 98 ¹H NMR (400 MHz, DMSO-d₆)δ 8.54 (s, 1H), 8.32 (s, 2H), 7.35-7.14 (m, 7H), 525.6 3.99-3.85 (m,7H), 3.80 (s, 2H), 3.72-3.63 (m, 4H), 2.98-2.89 (m, 2H), 1.95-1.80 (m,3H), 1.74-1.65 (m, 2H), 1.31-1.12 (m, 4H). 99 525.6 100 ¹H NMR (400 MHz,DMSO-d₆) δ 11.35 (b.s., 1H), 8.57 (s, 1H), 7.77 (d, J = 9.1 Hz, 525.61H), 7.64 (s, 1H), 7.49 (dd, J = 9.1, 2.6 Hz, 1H), 7.30-7.18 (m, 6H),7.18-7.12 (m, 1H), 3.97 (d, J = 5.7 Hz, 2H), 3.83-3.69 (m, 8H), 3.57 (s,2H), 2.84-2.75 (m, 2H), 2.16 (s, 3H), 1.97-1.82 (m, 2H), 1.82-1.68 (m,3H), 1.43-1.26 (m, 2H). 101 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H),8.35 (s, 2H), 7.43 (t, J = 8.1 Hz, 526.5 1H), 7.28 (m, 2H), 7.24-7.17(m, 3H), 3.96-3.89 (m, 10H), 3.86 (s, 2H), 3.73-3.65 (m, 4H). 102 ¹H NMR(400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.31 (s, 2H), 7.76 (d, J = 8.0 Hz,526.5 2H), 7.66 (d, J = 8.0 Hz, 2H), 7.21 (d, J = 13.8 Hz, 2H), 5.24 (s,2H), 3.92 (s, 6H), 3.86 (dd, J = 6.4, 3.8 Hz, 4H), 3.72-3.66 (m, 4H).103 ¹H NMR (400 MHz, Chloroform-d) δ 8.69 (s, 1H), 8.15 (s, 2H),7.34-7.25 (m, 527.4 4H), 7.16 (s, 1H), 4.99 (s, 2H), 4.04 (s, 3H), 3.98(m, 7H), 3.79 (m, 4H). 104 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H),8.31 (s, 2H), 7.35-7.14 (m, 7H), 527.6 4.13-4.04 (m, 2H), 3.93 (s, 3H),3.92-3.87 (m, 4H), 3.79 (s, 2H), 3.78-3.72 (m, 2H), 3.71-3.63 (m, 4H),3.56-3.42 (m, 1H), 2.92-2.85 (m, 1H), 2.75-2.61 (m, 2H), 2.60-2.51 (m,1H). 105 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.32 (s, 2H),7.35-7.14 (m, 7H), 527.6 4.13-4.04 (m, 2H), 3.93 (s, 3H), 3.93-3.87 (m,4H), 3.80 (s, 2H), 3.78-3.72 (m, 2H), 3.72-3.62 (m, 4H), 3.52-3.44 (m,1H), 2.92-2.85 (m, 1H), 2.74-2.63 (m, 2H), 2.60-2.51 (m, 1H). 106 ¹H NMR(400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.31 (s, 2H), 7.33-7.13 (m, 7H),527.6 4.19-4.04 (m, 2H), 3.93 (s, 3H), 3.92-3.87 (m, 4H), 3.79 (s, 2H),3.78-3.74 (m, 2H), 3.71-3.62 (m, 4H), 3.56-3.45 (m, 1H), 2.96-2.88 (m,1H), 2.78-2.64 (m, 2H), 2.63-2.54 (m, 1H). 107 1H NMR (400 MHz, DMSO-d6)δ 8.55 (s, 1H), 8.50 (s, 2H), 7.20 (d, J = 6.3 Hz, 528.4 2H), 4.18 (t, J= 6.4 Hz, 2H), 3.93 (d, J = 3.4 Hz, 7H), 3.77-3.65 (m, 4H), 2.54 (t, J =7.1 Hz, 2H), 2.46-2.40 (m, 4H), 2.01-1.89 (m, 2H), 1.73-1.63 (m, 4H).108 530.6 109 539.6 110 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 8.32(s, 2H), 7.35-7.15 (m, 7H), 539.6 4.24 (t, J = 5.6 Hz, 2H), 3.93 (s,3H), 3.92-3.86 (m, 4H), 3.80 (s, 2H), 3.71-3.66 (m, 2H), 3.63-3.55 (m,4H), 3.51 (t, J = 7.0 Hz, 2H), 2.21 (t, J = 8.1 Hz, 2H), 1.96-1.88 (m,2H). 111 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.31 (s, 2H),7.33-7.15 (m, 7H), 539.6 4.25 (t, J = 5.6 Hz, 2H), 3.92 (m, 7H), 3.80(s, 2H), 3.73-3.65 (m, 4H), 3.61 (t, J = 5.5 Hz, 2H), 3.51 (t, J = 7.0Hz, 2H), 2.21 (t, J = 8.1 Hz, 2H), 1.92 (p, J = 7.6 Hz, 2H). 112 ¹H NMR(400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.31 (s, 2H), 7.33-7.15 (m, 7H),539.7 3.97 (d, J = 5.9 Hz, 2H), 3.93 (s, 3H), 3.92-3.86 (m, 4H), 3.79(s, 2H), 3.71-3.62 (m, 4H), 2.81 (d, J = 10.9 Hz, 2H), 2.18 (s, 3H),1.99-1.85 (m, 2H), 1.82-1.68 (m, 3H), 1.43-1.28 (m, 2H). 113 ¹H NMR (400MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.31 (s, 2H), 7.34-7.14 (m, 7H), 539.73.99 (d, J = 6.5 Hz, 2H), 3.93 (s, 3H), 3.92-3.87 (m, 4H), 3.79 (s, 2H),3.71-3.62 (m, 4H), 2.87-2.78 (m, 1H), 2.68-2.58 (m, 1H), 2.15 (s, 3H),2.12-1.99 (m, 1H), 1.98-1.70 (m, 3H), 1.71-1.59 (m, 1H), 1.56-1.42 (m,1H), 1.19-0.99 (m, 1H). 114 539.7 115 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55(s, 1H), 8.32 (s, 2H), 7.35-7.15 (m, 7H), 539.7 4.00 (d, J = 6.4 Hz,2H), 3.94 (s, 3H), 3.93-3.88 (m, 4H), 3.80 (s, 2H), 3.71-3.65 (m, 4H),2.89-2.78 (m, 1H), 2.68-2.58 (m, 1H), 2.16 (s, 3H), 2.13-1.98 (m, 1H),1.96-1.72 (m, 3H), 1.71-1.42 (m, 2H), 1.18-1.00 (m, 1H). 116 ¹H NMR (400MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.32 (s, 2H), 7.34-7.15 (m, 7H), 540.74.22 (t, J = 5.9 Hz, 2H), 3.92 (s, 3H), 3.92-3.88 (m, 4H), 3.80 (s, 2H),3.73-3.64 (m, 4H), 2.74-2.64 (m, 6H), 2.45-2.37 (m, 4H). 117 541.6 118541.6 119 ¹H NMR (400 MHz, DMSO-d₆) δ 8.53 (s, 1H), 7.63 (b.s., 1H),7.29-7.10 (m, 543.6 7H), 4.18-4.05 (m, 2H), 3.93 (s, 3H), 3.84-3.73 (m,6H), 3.70-3.63 (m, 4H), 3.62-3.45 (m, 3H), 3.01-2.90 (m, 1H), 2.86-2.53(m, 3H). 120 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 7.62 (b.s., 1H),7.30-7.10 (m, 543.6 7H), 4.15-4.04 (m, 2H), 3.93 (s, 3H), 3.84-3.74 (m,6H), 3.70-3.63 (m, 4H), 3.56 (s, 2H), 3.50 (td, J = 11.1, 3.0 Hz, 1H),2.97-2.90 (m, 1H), 2.80-2.64 (m, 2H), 2.59 (dd, J = 12.3, 10.3 Hz, 1H).121 553.7 123 1H NMR (400 MHz, DMSO-d6) δ 8.73 (s, 2H), 8.56 (s, 1H),7.54 (d, J = 7.9 Hz, 553.7 1H), 7.47 (s, 1H), 7.44 (d, J = 7.8 Hz, 1H),7.33 (t, J = 7.6 Hz, 1H), 7.26 (d, J = 7.9 Hz, 1H), 7.22 (s, 2H), 7.15(d, J = 7.6 Hz, 1H), 4.17 (t, J = 6.5 Hz, 2H), 4.01 (m, 4H), 3.94 (s,3H), 3.74 (t, J = 5.1 Hz, 4H), 2.43-2.28 (m, 8H), 1.93 (m, 2H), 1.49 (m,4H), 1.37 (m, 2H). 124 554.7 125 ¹H NMR (400 MHz, DMSO-d₆) δ 8.53 (s,1H), 7.65 (s, 1H), 7.32-7.12 (m, 7H), 554.7 6.30 (s, 2H), 3.97 (d, J =5.8 Hz, 2H), 3.93 (s, 3H), 3.86-3.79 (m, 4H), 3.65 (s, 2H), 3.65-3.58(m, 4H), 2.84-2.73 (m, 2H), 2.16 (s, 3H), 1.93-1.82 (m, 2H), 1.80-1.70(m, 3H), 1.42-1.28 (m, 2H). 126 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s,1H), 8.32 (s, 2H), 7.37-7.14 (m, 7H), 554.7 4.23 (d, J = 6.0 Hz, 3H),3.95-3.88 (m, 7H), 3.80 (s, 2H), 3.72-3.65 (m, 4H), 2.81-2.73 (m, 2H),2.65-2.10 (m, 11H, buried under DMSO peak). 127 ¹H NMR (400 MHz,DMSO-d₆) δ 8.54 (s, 1H), 7.64 (s, 2H), 7.30-7.11 (m, 7H), 555.7 3.97 (d,J = 5.9 Hz, 2H), 3.93 (s, 2H), 3.81-3.73 (m, 4H), 3.72-3.64 (m, 4H),3.57 (s, 2H), 2.91-2.75 (m, 2H), 2.20 (s, 3H), 2.02-1.87 (m, 1H),1.82-1.70 (m, 4H), 1.43-1.27 (m, 2H). 128 555.7 129 555.7 130 557.6 131557.7 132 ¹H NMR (400 MHz, DMSO-d₆) δ 8.55 (s, 1H), 8.29 (s, 2H), 7.68(s, 1H), 7.23 (s, 558.6 1H), 7.19 (s, 1H), 6.99 (t, J = 8.8 Hz, 2H),6.87-6.73 (m, 2H), 3.99 (d, J = 5.9 Hz, 2H), 3.93 (s, 2H), 3.92-3.87 (m,4H), 3.74-3.67 (m, 4H), 2.84-2.75 (m, 2H), 2.16 (s, 3H), 1.95-1.83 (s,1H), 1.82-1.71 (m, 4H), 1.45-1.28 (m, 2H). 133 ¹H NMR (400 MHz, DMSO-d₆)δ 8.55 (s, 1H), 8.35 (s, 2H), 7.25-7.15 (m, 4H), 559.6 7.07-7.00 (m,3H), 4.01-3.96 (m, 2H), 3.96-3.91 (m, 7H), 3.76-3.68 (m, 4H), 2.85-2.74(m, 2H), 2.16 (s, 3H), 1.93-1.82 (m, 2H), 1.81-1.71 (m, 3H), 1.42-1.28(m, 2H). 134 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.31 (s, 2H),7.29 (m, 2H), 568.7 7.25-7.15 (m, 5H), 4.16 (t, J = 6.4 Hz, 2H), 3.91(m, 7H), 3.80 (s, 2H), 3.72-3.64 (m, 4H), 2.42 (t, J = 7.1 Hz, 2H),2.39-2.22 (m, 8H), 2.13 (s, 3H), 1.92 (t, J = 6.8 Hz, 2H). 135 ¹H NMR(400 MHz, DMSO-d₆) δ 8.52 (s, 1H), 7.65 (s, 1H), 7.34-7.10 (m, 7H),568.7 6.30 (s, 2H), 4.20 (q, J = 7.0 Hz, 2H), 3.97 (d, J = 5.9 Hz, 2H),3.86-3.79 (m, 4H), 3.65 (s, 2H), 3.64-3.59 (m, 4H), 2.85-2.75 (m, 2H),2.16 (s, 3H), 1.94-1.81 (m, 2H), 1.80-1.79 (m, 3H), 1.39 (t, J = 7.0 Hz,3H), 1.39-1.29 (m, 2H). 136 ¹H NMR (400 MHz, DMSO-d₆) δ 8.54 (s, 1H),8.32 (s, 2H), 7.37-7.12 (m, 7H), 568.7 4.14 (t, J = 6.4 Hz, 2H),3.95-3.87 (m, 7H), 3.80 (s, 2H), 3.71-3.63 (m, 4H), 2.43 (t, J = 7.2 Hz,2H), 2.43-2.15 (m, 8H), 2.10 (s, 3H), 1.97-1.85 (m, 2H). 137 1H NMR (400MHz, DMSO-d6) δ 8.55 (s, 1H), 8.28 (s, 2H), 7.59 (s, 2H), 7.21 (d, 568.7J = 4.2 Hz, 2H), 6.97 (d, J = 8.1 Hz, 2H), 6.77-6.68 (m, 2H), 4.17 (t, J= 6.4 Hz, 2H), 3.93 (s, 3H), 3.92-3.85 (m, 4H), 3.71 (t, J = 5.2 Hz,4H), 2.39 (t, J = 7.1 Hz, 2H), 2.33 (br.s, 4H), 2.17 (s, 3H), 1.93 (m,2H), 1.49 (m, 4H), 1.37 (m, 2H). 138 ¹H NMR (400 MHz, DMSO-d₆) δ 11.36(b.s., 1H), 8.53 (s, 1H), 7.64 (s, 1H), 569.7 7.30-7.10 (m, 7H), 4.20(q, J = 7.0 Hz, 2H), 3.96 (d, J = 5.9 Hz, 2H), 3.84-3.61 (m, 8H), 3.57(s, 2H), 2.84-2.73 (m, 2H), 2.16 (s, 3H), 1.93-1.82 (m, 2H), 1.80-1.69(m, 3H), 1.39 (t, J = 6.9 Hz, 3H), 1.40-1.27 (m, 2H). 139 ¹H NMR (400MHz, DMSO-d₆) δ 8.54 (s, 1H), 8.32 (s, 2H), 7.35-7.12 (m, 7H), 569.74.05-3.95 (m, 2H), 3.93 (s, 3H), 3.92-3.87 (m, 4H), 3.80 (s, 2H),3.71-3.64 (m, 4H), 3.56-3.46 (m, 2H), 3.05-2.89 (m, 2H), 2.05-1.90 (m,2H), 1.87-1.67 (m, 3H), 1.55-1.28 (m, 2H). 140 ¹H NMR (400 MHz, DMSO-d₆)δ 8.55 (s, 1H), 8.53 (s, 1H), 7.30-7.13 (m, 6H), 575.7 4.03-3.95 (m,6H), 3.93 (s, 3H), 3.78-3.69 (m, 4H), 2.84-2.75 (m, 2H), 2.16 (s, 3H),1.94-1.83 (m, 2H), 1.80-1.70 (m, 3H), 1.45-1.28 (m, 2H).

Biochemical Activity of Compounds

In order to assess the activity of chemical compounds against therelevant kinase of interest, the Caliper LifeSciences electrophoreticmobility shift technology platform is used. Fluorescently labeledsubstrate peptide is incubated in the presence of kinase and ATP so thata reflective proportion of the peptide is phosphorylated. At the end ofthe reaction, the mix of phosphorylated (product) and non-phosphorylated(substrate) peptides are passed through the microfluidic system of theCaliper EZ Reader 2, under an applied potential difference. The presenceof the phosphate group on the product peptide provides a difference inmass and charge between those of the substrate peptide, resulting in aseparation of the substrate and product pools in the sample. As thepools pass a LEDS within the instrument, these pools are detected andresolved as separate peaks. The ratio between these peaks thereforereflects the activity of the chemical matter at that concentration inthat well, under those conditions.

Kit wild type assay at Km: In each well of a 384-well plate, 0.2 ng/ulfinal (2 nM) of wild type Kit (Carna Bioscience 08-156) was incubated ina total of 12.5 ul of buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 10 mMMgCl2, 1 mM DTT) with 1 uM Srctide (5-FAM-GEEPLYWSFPAKKK-NH2) and 400 uMATP at 25 C for 90 minutes in the presence or absence of a dosedconcentration series of compound (1% DMSO final concentration). Thereaction was stopped by the addition of 70 ul of Stop buffer (100 mMHEPES pH 7.5, 0.015% Brij 35, 35 mM EDTA and 0.2% of Coating Reagent 3(Caliper Lifesciences)). The plate was then read on a Caliper EZReader 2(protocol settings: −1.9 psi, upstream voltage −700, downstream voltage−3000, post sample sip 35s). Data was normalized to 0% and 100%inhibition controls and the IC50 or EC50 calculated using a 4-parameterfit using GraphPad Prism.

Kit D816V assay at Km: In each well of a 384-well plate, 0.04 ng/ul (0.5nM) of D816V Kit (Carna Bioscience 08-156) was incubated in a total of12.5 ul of buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 10 mM MgCl2, 1mM DTT) with 1 uM Srctide (5-FAM-GEEPLYWSFPAKKK-NH2) and 15 uM ATP at 25C for 90 minutes in the presence or absence of a dosed concentrationseries of compound (1% DMSO final concentration). The reaction wasstopped by the addition of 70 ul of Stop buffer (100 mM HEPES pH 7.5,0.015% Brij 35, 35 mM EDTA and 0.2% of Coating Reagent 3 (CaliperLifesciences)). The plate was then read on a Caliper EZReader 2(protocol settings: −1.9 psi, upstream voltage −700, downstream voltage−3000, post sample sip 35s). Data was normalized to 0% and 100%inhibition controls and the IC50 or EC50 calculated using a 4-parameterfit using GraphPad Prism.

The Table below shows the activity of compounds described herein,against wild-type Kit and mutant Kit (the D816V mutant). In the Tablebelow, for D816V activity, the following designations are used: <1.00nM=A; 1.01-10.0 nM=B; 10.01-100.0 nM=C; and >100 nM=D. For wild-type Kitactivity, the following designations are used: <10 nM=A; 11-100 nM=B;100-1000 nM=C; >1000 nM=D.

INH-KITD816V INH-KITWT ENZ-KM ENZ-KM Compound IC50 (nM) IC50 (nM) IdStructure (Num) (Num) 1

C D 2

D D 3

D D 4

B C 5

C C 6

D D 7

C C 8

D D 9

D D 10

D D 11

B B 12

D D 13

B C 14

B B 15

C C 16

C C 17

D D 18

D D 19

D D 20

C D 21

D D 22

B B 23

B B 24

B C 25

C C 26

C C 27

D D 28

D D 29

A A 30

B C 31

B C 32

C C 33

C C 34

C C 35

A A 36

B B 37

B C 38

B B 39

B C 40

B C 41

D D 42

B C 43

B C 44

C C 45

B C 46

C C 47

C D 48

B B 49

C C 50

D D 51

C D 52

B B 53

C C 54

C C 55

C D 56

C D 57

A B 58

B B 59

B C 60

C C 61

C C 62

C D 63

C C 64

C D 65

C D 66

C D 67

D D 68

D D 69

C C 70

B C 71

B C 72

C 73

D D 74

C C 75

B C 76

C C 77

C D 78

C D 79

B B 80

B B 81

C D 82

D D 83

D 84

B B 85

B B 86

B C 87

B C 88

C C 89

A B 90

A B 91

B B 92

B C 93

B B 94

D D 95

B B 96

A B 97

A B 98

A B 99

B B 100

B C 101

C C 102

D D 103

D D 104

A B 105

A B 106

B B 107

C C 108

A A 109

A B 110

A B 111

A B 112

A B 113

A B 114

A B 115

B B 116

B B 117

A B 118

B B 119

A A 120

A A 121

A B 122

A A 123

C C 124

A A 125

A A 126

B B 127

A A 128

B B 129

B B 130

B A 131

A A 132

B B 133

B B 134

A A 135

A A 136

A B 137

B B 138

A A 139

B B 140

B B

The following compounds were at least 100 times more potent against theD816V mutant than against the wild-type Kit: 1; 18; 20; 47; 57; 62; 73;and 77.

The following compounds were at least 50 times, but less than 100 times,more potent against the D816V mutant than against the wild-type Kit: 46;75; 78; 89; 94; 96; 103; and 136.

The following compounds were at least 25 times, but less than 50 times,more potent against the D816V mutant than against the wild-type Kit: 4;5; 6; 7; 8; 11; 16; 22; 24; 25; 33; 36; 37; 39; 40; 43; 45; 48; 58; 64;66; 70; 71; 76; 79; 81; 86; 87; 90; 94; 95; 97; 99; 101; 106; 107; 108;109; 110; 111; 112; 113; 114; 116; 118; 121; 126; and 139.

The following compounds were at least 10 times, but less than 25 times,more potent against the D816V mutant than against the wild-type Kit: 9;10; 12; 13; 14; 15; 17; 26; 27; 29; 30; 31; 32; 35; 38; 42; 44; 49; 51;52; 53; 54; 55; 56; 59; 60; 61; 63; 65; 69; 74; 80; 82; 84; 85; 88; 91;98; 104; 105; 117; 120; 122; 123; 124; 131; 132; 133; 134; and 137.

The following compounds were at least 5 times, but less than 10 times,more potent against the D816V mutant than against the wild-type Kit: 3;34; 100; 102; 119; 125; 127; 128; 130; and 135.

The following compounds were less than 5 times more potent against theD816V mutant than against the wild-type Kit: 19; 21; 23; 28; 41k; 50;67; 68; 129; and 138.

Cellular Activity

HMC1.2 Proliferation Assay:

1000 HMC1.2 cells were incubated in 22 ul culture media (IMDM, 10% calfserum with Iron) in each well of a 384-well plate overnight in a tissueculture incubator (5% CO₂, 37° C.). A 10 point dose concentration seriesof compound (25 uM-1.5 nM) were then added to the cells as 3.1 ulcompound solution to each well (0.25% DMSO final concentration). After 3days incubation in tissue culture incubator, 25 ul CellTiter-Glo(Promega) solution was added to each well for ATP/cell viabilitymeasurement. Luminescence signal was obtained on Envision (Perkin Elmer)by US Lum 384 protocol. Data was normalized to 0% and 100% inhibitioncontrols and the IC50 was calculated using Four Parameter Logistic IC50curve fitting.

HMC1.2 Autophosphorylation Assay:

10,000 HMC1.2 cells were incubated in 22 ul culture media (phenol-redfree IMDM, no serum) in each well of a 384-well plate and serum starvedovernight in a tissue culture incubator (5% CO₂, 37° C.). A 10-pointdose concentration series of compound (25 uM-95.4 pM) were then added tothe cells in a volume of 3.1 ul to each well (0.25% DMSO finalconcentration). After 90 minutes, 6 ul of 5× AlphaLISA Lysis Buffer(Perkin Elmer) supplemented with a protease and phosphatase inhibitorcocktail (Cell Signaling Technologies) was added to each well and shakenat 450 rpm for 15 minutes at 4° C. 10 ul of phospho-Y719 c-Kit and totalc-Kit antibodies (15 nM final concentration, Cell SignalingTechnologies) and 50 ug/ml AlphaLISA rabbit acceptor beads (PerkinElmer) were added to each well and shaken at 300 rpm at room temperaturefor 2 hours. 10 ul of 100 ug/ml streptavidin donor beads (Perkin Elmer)were added to each well, blocked from light with aluminum adhesive andshaken at 300 rpm at room temperature for 2 hours. Fluorescence signalwas obtained on Envision (Perkin Elmer) by AlphaScreen 384 well HTSprotocol. Data was normalized to 0% and 100% inhibition controls and theIC50 was calculated using Four Parameter Logistic IC50 curve fitting.

The Table below shows the activity of compounds in a Mast cell leukemiacell line, HMC 1.2. This cell line contains Kit mutated at positionsV560G and D816V resulting in constitutive activation of the kinase. Thefollowing compounds were tested in an assay to measure direct inhibitionKit D816V kinase activity by assaying Kit autophosphorylation attyrosine 719 on the Kit protein. In addition, these compounds wereassayed for their ability to inhibit the dependence of these cells onKit kinase activity by disrupting their growth and proliferation.

In the Table below, the following designations are used: <10 nM=A;10.01-100 nM=B; 100.01-1000 nM=C; and 1000-10000 nM=D, >10000.01 nM=E.

HMC1.2 HMC1.2 Autophosphorylation Proliferation Compound IC50 IC50 1 E D5 D 7 D 13 C D 22 D 23 C D 30 D 1 E D 35 C C 36 C D 42 D 79 B D 84 C D85 C D 88 E 98 B D 99 C D 105 B C 107 D 108 B C 109 B D 111 C D 112 B D105 B C 119 D E 123 D D 125 A C 134 B C 136 C D 137 C D

INCORPORATION BY REFERENCE

All publications and patents mentioned herein are hereby incorporated byreference in their entirety as if each individual publication or patentwas specifically and individually indicated to be incorporated byreference.

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

We claim:
 1. A compound having structural formula I:

or a pharmaceutically acceptable salt or a tautomer thereof, wherein:each R⁴ is independently selected from —C(O)N(R²)(R²), —C₁-C₄ alkyl,—CN, —(C₀-C₄ alkylene)-N(R²)(R²), —O—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl),—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl), and —O—(C₀-C₄ alkylene)-(R¹), whereinR¹ is selected from —C₁-C₄ alkyl and heterocyclyl; each R² isindependently selected from hydrogen and unsubstituted C₁-C₄ alkyl eachR⁶ is independently selected from —C₁-C₄ alkyl or two R⁶ bound to thesame carbon atom are taken together to form oxo, or two R⁶ bound todifferent carbon atoms are taken together to form methylene,ethane-1,2-diyl, or propane-1,2-diyl forming a ring that is bridged tothe piperazine-1,4-diyl portion of the compound; each R⁸ isindependently selected from halo, —OH, —N(R²)(R²), C₁-C₄ alkyl, and—O—(C₁-C₄ alkyl); L is selected from a bond, —(C₁-C₄ alkylene)-, —O—,—S—, —SO₂—, —N(R²)—, —O—(C₁-C₄ alkylene)-, —(C₁-C₄ alkylene)-O—, —(C₁-C₄alkylene)-N(R²)—, —N(R²)—(C₁-C₄ alkylene)-, —N(R²)—CO—(C₁-C₄ alkylene)-,—CO—N(R²)—(C₁-C₄ alkylene)-; n is 0, 1, 2, or 3; m is 0, 1, 2, 3, or 4;p is 0, 1, or 2; q is 0, 1, 2, or 3; ring A is monocyclic or bicyclicaryl, heteroaryl, carbocyclyl or heterocyclyl; each R¹⁰ is independentlyselected from halo, —OH, —CN, —C(O)N(R²)(R²), C₁-C₄ alkyl, —O—(C₁-C₄alkyl), and heterocyclyl, or two R¹° bound to adjacent ring carbon atomsare taken together to form methylenedioxy; wherein unless otherwisespecified any alkyl, or alkylene portion of the compound is optionallysubstituted; and wherein the compound is not


2. The compound of claim 1, wherein any alkyl, or alkylene portion ofthe compound is optionally and independently substituted with one ormore substitutents independently selected from halo, —OH, C₁-C₄ alkyl,C₁-C₄ haloalkyl, —O—(C₁-C₄ alkyl), and ═O; and any heterocyclyl portionof the compound is optionally and independently substituted with one ormore substitutents independently selected from halo, —OH, C₁-C₄ alkyl,C₁-C₄ haloalkyl, —O—(C₁-C₄ alkyl), and ═O.
 3. The compound of claim 1,wherein: each R⁴ is independently selected from —C(O)N(R²)(R²),—O—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl), and —O—(C₀-C₄ alkylene)-(R¹),wherein: R¹ is selected from —C₁-C₄ alkyl, and a saturated heterocyclyl;each R² is independently selected from hydrogen and methyl; each alkylor alkylene is optionally substituted with one or more substituentsindependently selected from halo, —OH and —O—(C₁-C₃ alkyl); thesaturated heterocyclyl is optionally substituted on a substitutable ringnitrogen with methyl; and the saturated heterocyclyl is optionallysubstituted on a substitutable ring carbon with with one or moresubstituents independently selected from methyl, halo, —OH.
 4. Thecompound of claim 3, wherein each R⁴ is independently selected fromC(O)NH₂, —OCF₂, —OCH₂CH₃, —OCH₃, 1-methyl-4-fluoropiperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-yloxy,1-(2-hydroxyethyl)-pyrrolidin-3-yloxy, 1-methylazetidin-3-ylmethoxy,1-methylpiperidin-3-ylethoxy, 1-methylpiperidin-3-ylmethoxy,1-methylpiperidin-4-ylmethoxy, 1-methylpiperidin-4-yloxy,1-methylpyrrolidin-3-ylmethoxy, 1-methylpyrrolidin-3-yloxy,2,3-dihydroxypropoxy, 2-oxopiperidin-4-ylmethoxy,2-oxopyrrolidin-1-ylethoxy, 3-methyloxetan-3-ylmethoxy,4-methylmorpholin-2-ylmethoxy, 4-methylpiperazin-1-ylethoxy,4-methylpiperazin-1-ylpropoxy, azetidin-3-ylmethoxy, methoxyethoxy,morpholin-2-ylmethoxy, piperidin-1-ylpropoxy, piperidin-3-ylmethoxy,piperidin-4-ylmethoxy, piperidin-4-yloxy, pyrrolidin-1-ylpropoxy,pyrrolidin-3-ylmethoxy, and pyrrolidin-3-yloxy.
 5. The compound of claim4, wherein each R⁴ is independently selected from OCH₂CH₃, OCH₃,1-(2-hydroxyethyl)-piperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-yloxy,1-(2-hydroxyethyl)-pyrrolidin-3-yloxy,1-methyl-4-fluoropiperidin-4-ylmethoxy, 1-methylazetidin-3-ylmethoxy,1-methylpiperidin-3-ylethoxy, 1-methylpiperidin-3-ylmethoxy,1-methylpiperidin-4-ylmethoxy, 1-methylpiperidin-4-yloxy,1-methylpyrrolidin-3-ylmethoxy, 1-methylpyrrolidin-3-yloxy,2,3-dihydroxypropoxy, 2-oxopiperidin-4-ylmethoxy,2-oxopyrrolidin-1-yleithoxy, 3-methyloxetan-3-ylmethoxy,4-methylmorpholin-2-ylmethoxy, 4-methylpiperazin-1-ylethoxy,4-methylpiperazin-1-ylpropoxy, azetidin-3-ylmethoxy, methoxyethoxy,morpholin-2-ylmethoxy, piperidin-1-ylpropoxy, piperidin-3-ylmethoxy,piperidin-4-ylmethoxy, piperidin-4-yloxy, pyrrolidin-3-ylmethoxy, andpyrrolidin-3-yloxy.
 6. The compound of claim 1, wherein each R⁶ isindependently selected from C₁-C₄ alkyl, and ═O, or two R⁶ onnon-adjacent carbon ring atoms are taken together to formethane-1,2-diyl, propane-1,3-diyl or butane-1,4-diyl thereby forming aring that is bridged to the piperazine-1,4-diyl portion of the compound.7. The compound of claim 6, wherein each R⁶ is independently methyl or═O, or two R⁶ non-adjacent carbon ring atoms are taken together to forman ethane-1,2-diyl thereby forming a ring that is bridged to thepiperazine-1,4-diyl portion of the compound.
 8. The compound of claim 1,wherein each R⁸ is independently selected from ═O and —NH₂.
 9. Thecompound of claim 1, wherein each R¹° is independently selected from—OH, —OCH₃, —F, —CH₃, —CN, —C(O)NH₂, —OCF₂, —Cl, —CF₃, —OCF₃, andt-butyl, or two R¹° are taken together to form methylenedioxy.
 10. Thecompound of claim 9, wherein each R¹° is independently selected from OH,—OCH₃, —F, —CH₃, —CN, —C(O)NH₂, —OCF₂, and —Cl.
 11. The compound ofclaim 1, wherein L is selected from a bond, —CH₂—, —CH₂CH₂—, —NH—, 0, S,CH₂O—*, —OCH₂—*, —OCH(CH₃)—*, —N(CH₃)CH₂—*, —NHCH₂—*, —NHC(O)CH₂—*,—C(O)NH—*, —NHCH(CH₃)—*, and —SO₂—, wherein “*” represents a portion ofL bound to ring A.
 12. The compound of claim 11, wherein L is selectedfrom —CH₂—, —CH₂CH₂—, —NH—, —O—, —S—, —CH₂O—*, —N(CH₃)CH₂—*,—OCH(CH₃)—*, and —OCH(CH₃)—*.
 13. The compound of claim 1, wherein ringA is selected from phenyl, thiophenyl, indolinyl,1,2,3,4-tetrahydroquinoline, pyridinyl, thiophenyl, and C₃-C₆cycloalkyl.
 14. The compound of claim 13, wherein ring A is selectedfrom phenyl, and thiophen-2-yl.
 15. A compound having the structuralformula II:

or a pharmaceutically acceptable salt or tautomer thereof, wherein: oneof R^(4a) or R^(4b) is selected from hydrogen, —O—CH₃, —O—CH₂CH₃ and—O—CH₂CH₂—O—CH₃; the other of R^(4a) or R^(4b) is selected from —O—CH₃,—O—CH₂CH₃, —O—CH₂CH₂—O—CH₃ and —(C₀-C₄ alkylene)-(saturatedheterocyclyl), wherein the saturated heterocyclyl is optionallysubstituted on a substitutable ring nitrogen with methyl; and thesaturated heterocyclyl is optionally substituted on a substitutable ringcarbon with one or more substituents independently selected from methyl,halo, and —OH; R^(6a) is hydrogen; R^(6b) is selected from hydrogen, ═O,and —CH₃; or R^(6a) and R^(6b) are taken together with the carbon atomsto which they are bound to form ethane-1,2-diyl thereby forming a ringbridged to the piperazine-1,4-diyl portion of the compound; R^(8a) isselected from hydrogen, —NH₂ and —OH; L is selected from —CH₂—,—CH₂CH₂—, —NH—, —O—, —S—, —CH₂O—*, —N(CH₃)CH₂—*, —OCH(CH₃)—*, and—OCH(CH₃)—*; R^(10a) is selected from hydrogen, —OCH₃, —C(O)NH₂ andhalo; and R^(10b) is selected from hydrogen, —OCH₃, —OCF₂, halo, —CH₃,CN, and OH.
 16. The compound of claim 15, wherein: one of R^(4a) orR^(4b) is selected from hydrogen, —O—CH₃, —O—CH₂CH₃ and —O—CH₂CH₂—O—CH₃;the other of R^(4a) or R^(4b) is selected from —O—CH₃, —O—CH₂CH₃,—O—CH₂CH₂—O—CH₃, 1-(2-hydroxyethyl)-piperidin-4-ylmethoxy,1-(2-hydroxyethyl)-piperidin-4-yloxy,1-(2-hydroxyethyl)-pyrrolidin-3-yloxy,1-methyl-4-fluoropiperidin-4-ylmethoxy, 1-methylazetidin-3-ylmethoxy,1-methylpiperidin-3-ylethoxy, 1-methylpiperidin-3-ylmethoxy,1-methylpiperidin-4-ylmethoxy, 1-methylpiperidin-4-yloxy,1-methylpyrrolidin-3-ylmethoxy, 1-methylpyrrolidin-3-yloxy,2,3-dihydroxypropoxy, 2-oxopiperidin-4-ylmethoxy,2-oxopyrrolidin-1-yleithoxy, 3-methyloxetan-3-ylmethoxy,4-methylmorpholin-2-ylmethoxy, 4-methylpiperazin-1-ylethoxy,4-methylpiperazin-1-ylpropoxy, azetidin-3-ylmethoxy,morpholin-2-ylmethoxy, piperidin-1-ylpropoxy, piperidin-3-ylmethoxy,piperidin-4-ylmethoxy, piperidin-4-yloxy, pyrrolidin-3-ylmethoxy, andpyrrolidin-3-yloxy; R^(10a) is selected from hydrogen, —OCH₃, andfluoro; and R^(10b) is selected from hydrogen, —OCH₃, —OCF₂, fluoro,chloro, —CH₃, CN, OH, and —C(O)NH₂.
 17. A pharmaceutical compositioncomprising a compound of claim 1 and a pharmaceutically acceptablecarrier.
 18. A method of treating mastocytosis comprising administeringto a patient a therapeutically effective amount of a compound of claim 1or a pharmaceutical composition of claim
 17. 19. A method of of treatinggastrointestinal stromal tumor, the method comprising administering to apatient a therapeutically effective amount of a compound of claim 1 or apharmaceutical composition of claim
 17. 20. A method of treating acondition mediated by mutant Kit, wherein said Kit is mutated at residueD816, the method comprising administering to a patient a therapeuticallyeffective amount of a compound of claim 1 or a pharmaceuticalcomposition of claim
 17. 21. A compound having structural formula III:

or a pharmaceutically acceptable salt or tautomer thereof, wherein eachof X, Y, and Z is independently, C or N, each of which may be attachedto one or more R⁴; each R⁴ is independently selected from—C(O)N(R²)(R²), —C₁-C₄ alkyl, —CN, —(C₀-C₄ alkylene)-N(R²)(R²),—O—(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl), —(C₁-C₄ alkylene)-O—(C₁-C₃ alkyl),and —O—(C₀-C₄ alkylene)-(R¹), wherein R¹ is selected from —C₁-C₄ alkyland heterocyclyl; each R² is independently selected from hydrogen andunsubstituted C₁-C₄ alkyl; each R⁶ is independently selected from —C₁-C₄alkyl or two R⁶ bound to the same carbon atom are taken together to form═O, or two R⁶ bound to different carbon atoms are taken together to formmethylene, ethane-1,2-diyl or propane-1,3-diyl thereby forming a ringthat is bridged to the piperazine-1,4-diyl portion of the compound; eachR⁸ is independently selected from halo, —OH, —N(R²)(R²), C₁-C₄ alkyl,and —O—(C₁-C₄ alkyl); L is selected from a bond, —(C₁-C₄ alkylene)-,—O—, —S—, —SO₂—, —N(R²)—, —O—(C₁-C₄ alkylene)-, —(C₁-C₄ alkylene)-O—,—(C₁-C₄ alkylene)-N(R²)—, —N(R²)—(C₁-C₄ alkylene)-, —N(R²)—CO—(C₁-C₄alkylene)-, —CO—N(R²)—(C₁-C₄ alkylene)-; n is 0, 1, 2, or 3; m is 0, 1,2, 3, or 4; p is 0, 1, or 2; q is 0, 1, 2, or 3; ring A is monocyclic orbicyclic aryl, heteroaryl, carbocyclyl or heterocyclyl; each R¹⁰ isindependently selected from halo; —OH, —CN, —C(O)N(R²)(R²), C₁-C₄ alkyl,—O—(C₁-C₄ alkyl), or heterocyclyl, or two R¹° bound to adjacent ringcarbon atoms are taken together to form methylenedioxy; and whereinunless otherwise specified any alkyl, or alkylene portion of thecompound is optionally substituted.
 22. A pharmaceutical compositioncomprising a compound of claim
 21. 23. A method of treating mastocytosiscomprising administering to a patient a therapeutically effective amountof a compound of claim 21 or a pharmaceutical composition of claim 22.24. A method of of treating gastrointestinal stromal tumor, the methodcomprising administering to a patient a therapeutically effective amountof a compound of claim 21 or a pharmaceutical composition of claim 22.25. A method of treating a condition mediated by mutant Kit, whereinsaid Kit is mutated at residue D816, the method comprising administeringto a patient a therapeutically effective amount of a compound of claim21 or a pharmaceutical composition of claim
 22. 26. A compound that isat least 5 times, but less than 10 times, more active against the D816Vmutation of Kit than against wild-type Kit when measured in abiochemical assay.
 27. A compound that is at least 10 times, but lessthan 25 times, more active against the D816V mutation of Kit thanagainst wild-type Kit when measured in a biochemical assay.
 28. Acompound that is at least 25 times, but less than 50 times, more activeagainst the D816V mutation of Kit than against wild-type Kit whenmeasured in a biochemical assay.
 29. A compound that is at least 50times, but less than 100 times, more active against the D816V mutationof Kit than against wild-type Kit when measured in a biochemical assay.30. A compound that is at least 100 times more active against the D816Vmutation of Kit than against wild-type Kit when measured in abiochemical assay.